Chemiluminescence analyzer

Abstract

The present disclosure relates to a chemiluminescence analyzer, comprising: a sample preparation device configured to pre-dilute a blood sample under test by using a diluent, and mix the pre-diluted blood sample under test and a reaction reagent in a reaction vessel to obtain a sample liquid under test; a substrate providing device configured to provide a luminescent substrate into the reaction vessel containing the sample liquid under test so as to carry out a chemiluminescence reaction; a photometric device, at least comprising a photometer; and a data processing device configured to obtain a test result of human chorionic gonadotropin (HCG) in the blood sample under test on the basis of a luminescent signal and a preset calibration model. A first linear test range of the photometer is [A1, A2] photon count / second, and in the first linear test range, the intensity of the luminescent signal generated in the chemiluminescence reaction is linearly related to the photon count output by the photometer, wherein A1 is less than or equal to 2000 and A2 is greater than or equal to 108. Therefore, the testing efficiency and accuracy of HCG can be improved without increasing the costs.

Description

Chemiluminescence analyzer Technical Field

[0001] The present disclosure relates to the field of chemiluminescence detection, and in particular to a chemiluminescence analyzer. Background Art

[0002] Chemiluminescence immunoassay (CLIA) is an immunoassay technique that combines a chemiluminescence system with an immune response and is used to detect antigens, antibodies, hormones, fatty acids, vitamins, and drugs. Chemiluminescence occurs when a specific substance absorbs some chemical energy during a chemical reaction, reaching an excited state. Upon returning to the ground state, the energy released is released as photons, resulting in luminescence. CLIA combines the high sensitivity of chemiluminescence with the simplicity and rapid response of enzyme-linked immunosorbent assays (ELISAs), making standardized experimental procedures easy to use. It is now widely used in biological and medical research and clinical diagnosis.

[0003] However, for chemiluminescence analyzers based on chemiluminescence immunoassay, it is difficult to achieve both detection sensitivity and linear detection range, especially for analytes with a wide range, such as human chorionic gonadotropin (HCG).

[0004] Human chorionic gonadotropin (HCG) is a glycoprotein secreted primarily by placental trophoblast cells. HCG is composed of α and β subunits, each containing 237 amino acids and a molecular weight of approximately 38,000 daltons. Each subunit is composed of a single polypeptide: the α subunit consists of 92 amino acids with a molecular weight of approximately 14,900 daltons, and the β subunit consists of 145 amino acids with a molecular weight of approximately 23,000 daltons. HCG's structure is 70% protein and 30% sugar.

[0005] HCG can be found in urine, blood, and placenta. Each matrix contains different forms of HCG, but in the blood it primarily exists as intact HCG. The α subunit is structurally similar to the α subunits of follicle-stimulating hormone, luteinizing hormone, and thyroid-stimulating hormone secreted by the pituitary gland; the β subunits differ in structure, and these differences in β subunits contribute to their respective immunological and biological specificities.

[0006] The primary function of human chorionic gonadotropin (hCG) is to stimulate the corpus luteum, promoting the sustained secretion of estrogen and progesterone, thereby promoting the formation of the uterine decidua and the maturation of the placenta. HCG is produced by trophoblast transitional cells and syncytial cells, regulated by multiple factors, including autocrine and paracrine factors. In a normal pregnancy, hCG production begins two to eight days after conception. In early pregnancy, hCG levels double in approximately 31-48 hours, reaching a peak around 11 weeks of gestation. Levels then slowly decrease until the 18th to 20th week of gestation, where they remain relatively constant. HCG testing is crucial for early pregnancy diagnosis and monitoring. In early pregnancy, a 35–50% increase in hCG levels within two days indicates a possible ectopic pregnancy. Abnormally low or rapidly decreasing hCG levels indicate a high risk of ectopic pregnancy. On the other hand, elevated hCG levels in mid-pregnancy can indicate adverse pregnancy outcomes, such as gestational hypertension or fetal growth restriction.

[0007] The concentration distribution of HCG is relatively wide in normal women who are not pregnant, pregnant, and those with trophoblastic cell diseases: for pregnant people, their HCG level can increase from a few hundred mIU / mL in early pregnancy to a maximum of about 200,000 mIU / mL, and then decrease to a level within about 10,000 mIU / mL; for patients with trophoblastic cell diseases (including choriocarcinoma), their HCG concentration can be as high as 1,000,000 mIU / mL or even higher; and for non-pregnant women, their HCG concentration is usually below 5 mIU / mL, and some can even be as low as 0.5 mIU / mL.

[0008] That is, HCG is secreted by placental trophoblast cells and its content in non-pregnant people is very low, less than 5mIU / ml. Therefore, high detection sensitivity is the key performance to ensure that HCG test results can be used clinically to assist in determining pregnancy. At the same time, in the pregnancy monitoring scenario, it is necessary to continuously measure the HCG concentration in the early stages of pregnancy (approximately 200,000 mIU / ml). Therefore, a wide linear detection range is the key performance to ensure that HCG test results can be used clinically to monitor pregnancy status.

[0009] Therefore, for chemiluminescence analyzers, in the HCG detection scenario, the linear range that needs to be accurately detected spans 6 to 7 orders of magnitude. However, the current chemiluminescence analyzers and their supporting reagents can only achieve a linear detection upper limit of no more than 15,000 mIU / mL, which is far from covering routine pregnancy test scenarios. Summary of the Invention

[0010] Against this backdrop, the present invention aims to provide a chemiluminescence analyzer based on chemiluminescence immunoassay (CLI) with a wide linear detection range, balancing sensitivity with an upper limit of linear detection. In particular, for samples with high concentrations of the analyte (e.g., HCG or β-HCG), dilution and re-testing are not required, improving efficiency and saving reagent costs.

[0011] A first aspect of the present disclosure provides a chemiluminescence analyzer, comprising:

[0012] A sample preparation device is at least used to: for a blood sample to be tested for human chorionic gonadotropin, before the blood sample is tested for human chorionic gonadotropin, pre-dilute the blood sample with a diluent at a preset dilution ratio, and mix the pre-diluted blood sample with a reaction reagent in a reaction container so that human chorionic gonadotropin in the pre-diluted blood sample reacts with the reaction reagent to obtain a sample solution to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent for human chorionic gonadotropin antibodies with a marker;

[0013] The substrate providing device is used to provide the luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested and the luminescent substrate can undergo chemiluminescent reaction;

[0014] A photometric device, the photometric device at least includes a photometer, the photometer is used to detect the luminescent signal generated in the chemiluminescent reaction, wherein the first linear detection range of the photometer is [A1, A2] photon counts / second, the first linear detection range represents the range of photon counts output by the photometer, within the first linear detection range, the intensity of the luminescent signal generated in the chemiluminescent reaction is linearly related to the photon counts output by the photometer, wherein A1 is less than or equal to 2000 and A2 is greater than or equal to 10 8 ;

[0015] The data processing device is used to obtain the detection result of human chorionic gonadotropin in the blood sample to be tested based on the luminescent signal and the preset calibration model.

[0016] A second aspect of the present disclosure provides a chemiluminescence analyzer, comprising:

[0017] A sample preparation device, used for preparing a sample liquid to be tested from a blood sample to be tested;

[0018] A substrate providing device, used for providing a luminescent substrate;

[0019] A photometric device, the photometric device at least includes a photometer, the photometer is used to detect the luminescent signal generated in the chemiluminescent reaction, wherein the first linear detection range of the photometer is [A1, A2] photon counts / second, the first linear detection range represents the range of photon counts output by the photometer, within the first linear detection range, the intensity of the luminescent signal generated in the chemiluminescent reaction is linearly related to the photon counts output by the photometer, wherein A1 is less than or equal to 2000 and A2 is greater than or equal to 10 8 ;

[0020] controller, configured as

[0021] Obtain the test items for the blood sample to be tested,

[0022] When the test item is human chorionic gonadotropin test item:

[0023] The sample preparation device is controlled to pre-dilute the blood sample to be tested with a diluent, and the pre-diluted blood sample to be tested is mixed with a reaction reagent in a reaction container, so that the human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample liquid to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent with human chorionic gonadotropin antibodies with a marker. The substrate providing device is controlled to provide a luminescent substrate to the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested reacts with the luminescent substrate to undergo a chemiluminescent reaction.

[0024] Control the luminometer to detect the luminescent signal generated in the chemiluminescent reaction,

[0025] The detection result of human chorionic gonadotropin in the blood sample to be tested is obtained according to the luminescent signal and the preset calibration model.

[0026] A third aspect of the present disclosure provides a chemiluminescence analyzer, comprising:

[0027] A sample preparation device is at least used to: pre-dilute a blood sample to be tested for human chorionic gonadotropin testing using a diluent at a preset dilution ratio before performing a human chorionic gonadotropin test, and mix the pre-diluted blood sample to be tested with a reaction reagent in a reaction container so that human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent for human chorionic gonadotropin antibodies with a marker;

[0028] The substrate providing device is used to provide a luminescent substrate into a reaction container containing a sample liquid to be tested, so that the marker in the sample liquid to be tested and the luminescent substrate undergo a chemiluminescent reaction, wherein the second linear detection range of the luminescent substrate is [B1, B2] photon counts / second, and the second linear detection range represents the range of the intensity of the luminescent signal generated in the chemiluminescent reaction. Within the second linear detection range, the number of markers participating in the chemiluminescent reaction and the intensity of the luminescent signal generated in the chemiluminescent reaction are linearly related, wherein B1 is less than or equal to 3000 and B2 is greater than or equal to 10 8 , and / or, the ratio of B2 to B1 (B2 / B1) is greater than or equal to 30,000;

[0029] A photometric device, the photometric device comprising at least a photometer, the photometer being used to detect the luminescent signal generated in the chemiluminescent reaction;

[0030] The data processing device is used to obtain the detection result of the human chorionic gonadotropin in the blood sample to be tested according to the luminescent signal and the preset calibration model.

[0031] A fourth aspect of the present disclosure provides a chemiluminescence analyzer, comprising:

[0032] A sample preparation device, used for preparing a sample liquid to be tested from a blood sample to be tested;

[0033] A substrate providing device, used for providing a luminescent substrate;

[0034] A photometric device, the photometric device comprising at least a photometer, the photometer being used to detect the luminescent signal generated in the chemiluminescent reaction;

[0035] controller, configured as

[0036] Obtain the test items for the blood sample to be tested,

[0037] When the test item is human chorionic gonadotropin test item:

[0038] The sample preparation device is controlled to pre-dilute the blood sample to be tested with a diluent, and the pre-diluted blood sample to be tested is mixed with a reaction reagent in a reaction container, so that the human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample liquid to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent with a marker for human chorionic gonadotropin antibodies.

[0039] The substrate providing device is controlled to provide the luminescent substrate to the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested reacts with the luminescent substrate to perform chemiluminescent reaction.

[0040] Control the luminometer to detect the luminescent signal generated in the chemiluminescent reaction,

[0041] The detection result of human chorionic gonadotropin in the blood sample to be tested is obtained according to the luminescence signal and the preset calibration model.

[0042] The second linear detection range of the luminescent substrate is [B1, B2] photon counts / second, and the second linear detection range represents the range of the intensity of the luminescent signal generated in the chemiluminescent reaction. Within the second linear detection range, the number of markers participating in the chemiluminescent reaction is linearly related to the intensity of the luminescent signal generated in the chemiluminescent reaction, wherein B1 is less than or equal to 3000 and B2 is greater than or equal to 10 8 , and / or, the ratio of B2 to B1 B2 / B1 is greater than or equal to 30000.

[0043] A fifth aspect of the present disclosure is a chemiluminescence analyzer, characterized by comprising:

[0044] A sample preparation device, configured to pre-dilute all blood samples to be tested containing human chorionic gonadotropin as a test substance using a diluent, and to mix the pre-diluted blood samples to be tested with a reaction reagent in a reaction container so that the human chorionic gonadotropin in the pre-diluted blood samples to be tested reacts with the reaction reagent to obtain a sample solution to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent containing human chorionic gonadotropin antibodies with a marker;

[0045] The substrate providing device is used to provide the luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested and the luminescent substrate can undergo chemiluminescent reaction;

[0046] A photometric device, the photometric device at least includes a photometer, the photometer is used to detect the luminescence signal generated in the chemiluminescence reaction, the photometer includes a receiving component and a processing component, the receiving component is configured to receive the light signal generated in the chemiluminescence reaction and convert the light signal into a corresponding electrical signal, the processing component is configured to be electrically connected to the receiving component and receive the electrical signal from the receiving component, the processing component includes a first photon counting module and a second photon counting module, the first photon counting module is configured to detect the number of pulses of the electrical signal using a pulse recognition method to obtain a first photon counting result, the second photon counting module is configured to process the electrical signal to obtain a parameter representing the number of photons in the light signal, and obtain a second photon counting result based on the parameter representing the number of photons in the light signal and a preset calibration function, wherein the calibration function represents a mapping relationship between the parameter representing the number of photons in the light signal and the photon counting result, and the processing component is further configured to obtain a photon count output by the photometer based on the first photon counting result and the second photon counting result, as well as the output photon count;

[0047] The data processing device is used to obtain the detection result of human chorionic gonadotropin in the blood sample to be tested based on the luminescent signal and the preset calibration model.

[0048] A sixth aspect of the present disclosure provides a chemiluminescence analyzer, comprising:

[0049] A sample preparation device, used for preparing a sample liquid to be tested from a blood sample to be tested;

[0050] A substrate providing device, used for providing a luminescent substrate;

[0051] A photometric device, the photometric device at least includes a photometer, the photometer is used to detect the luminescence signal generated in the chemiluminescence reaction, the photometer includes a receiving component and a processing component, the receiving component is configured to receive the light signal generated in the chemiluminescence reaction and convert the light signal into a corresponding electrical signal, the processing component is configured to be electrically connected to the receiving component and receive the electrical signal from the receiving component, the processing component includes a first photon counting module and a second photon counting module, the first photon counting module is configured to detect the number of pulses of the electrical signal using a pulse recognition method to obtain a first photon counting result, the second photon counting module is configured to process the electrical signal to obtain a parameter representing the number of photons in the light signal, and obtain a second photon counting result based on the parameter representing the number of photons in the light signal and a preset calibration function, wherein the calibration function represents a mapping relationship between the parameter representing the number of photons in the light signal and the photon counting result, and the processing component is further configured to obtain a photon count output by the photometer based on the first photon counting result and the second photon counting result, as well as the output photon count;

[0052] controller, configured as

[0053] Obtain the test items for the blood sample to be tested,

[0054] When the test item is human chorionic gonadotropin test item:

[0055] The sample preparation device is controlled to pre-dilute the blood sample to be tested with a diluent, and the pre-diluted blood sample to be tested is mixed with a reaction reagent in a reaction container, so that the human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample liquid to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent with a marker for human chorionic gonadotropin antibodies.

[0056] The substrate providing device is controlled to provide the luminescent substrate to the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested reacts with the luminescent substrate to perform chemiluminescent reaction.

[0057] Control the luminometer to detect the luminescent signal generated in the chemiluminescent reaction,

[0058] The detection result of human chorionic gonadotropin in the blood sample to be tested is obtained according to the luminescent signal and the preset calibration model.

[0059] A seventh aspect of the present disclosure provides a chemiluminescence analyzer, comprising:

[0060] HCG measuring unit, used to obtain the test result of human chorionic gonadotropin of the blood sample to be tested;

[0061] a mode selection unit, configured to select a first HCG detection mode or a second HCG detection mode; and

[0062] Controller, configured as,

[0063] Mode selection of the receiving mode selection unit,

[0064] When the mode selection unit selects the first HCG detection mode, the HCG measurement unit is controlled to perform a first measurement process, wherein the first measurement process includes: pre-diluting at least a portion of the blood sample to be tested using a diluent, and performing chemiluminescence measurement on the pre-diluted portion of the blood sample to be tested to obtain and output a first HCG detection result; and

[0065] When the mode selection unit selects the second HCG detection mode, the HCG measurement unit is controlled to execute a second measurement process, which includes: performing chemiluminescence measurement on an undiluted portion of the blood sample to be tested to obtain a second HCG detection result; when the second HCG detection result is abnormal, pre-diluting another portion of the blood sample to be tested with a diluent, and performing chemiluminescence measurement on the pre-diluted portion of the blood sample to be tested to obtain and output a third HCG detection result.

[0066] An eighth aspect of the present disclosure provides a chemiluminescence analyzer, comprising:

[0067] A sample preparation device, configured to mix a blood sample to be tested with a reaction reagent in a reaction container so that a substance to be tested in the blood sample to be tested reacts with the reaction reagent to obtain a sample liquid to be tested, wherein the reaction reagent includes a capture reagent and a detection reagent with a marker;

[0068] A reagent carrying device, the reagent carrying device is used to place the test kit, the test kit carries the reaction reagent corresponding to the test item, and the test kit corresponding to the human chorionic gonadotropin test item also carries a diluent;

[0069] A substrate providing device, configured to provide a luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested reacts with the luminescent substrate to undergo a chemiluminescent reaction;

[0070] A photometric device, the photometric device comprising at least a photometer, the photometer being used to detect the luminescent signal generated in the chemiluminescent reaction;

[0071] a data processing device, configured to obtain a detection result of the substance to be detected in the blood sample to be tested based on the luminescence signal and a preset calibration model;

[0072] The control device is used to control the sample preparation device to pre-dilute the blood sample to be tested with a diluent in a kit corresponding to the human chorionic gonadotropin test item at a preset dilution ratio when the test item of the blood sample to be tested is the human chorionic gonadotropin test item, and to mix the pre-diluted blood sample to be tested with at least part of the reaction reagent in the kit corresponding to the human chorionic gonadotropin test item in the reaction container, so that the human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample liquid to be tested.

[0073] Controlling the substrate providing device to provide a luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested reacts with the luminescent substrate to undergo a chemiluminescent reaction.

[0074] The photometer is controlled to detect the luminescent signal generated in the chemiluminescent reaction, and the detection result of the human chorionic gonadotropin in the blood sample to be tested is obtained according to the luminescent signal and a preset calibration model.

[0075] The chemiluminescence analyzer provided in various aspects of the present disclosure can achieve efficient and accurate determination of HCG concentration without increasing the detection cost through the design of the photometer and pre-dilution of the blood sample to be tested. BRIEF DESCRIPTION OF THE DRAWINGS

[0076] The present disclosure will be more clearly described below with reference to the following examples and accompanying drawings. The above advantages and other advantages will become clear to those skilled in the art through the detailed description of the embodiments of the present disclosure. The accompanying drawings are only used to illustrate preferred embodiments and should not be considered as limiting the present disclosure. In the accompanying drawings:

[0077] FIG1 shows a schematic block diagram of a chemiluminescence analyzer provided in an embodiment of the present disclosure.

[0078] FIG2 and FIG3 show a calibration curve of a chemiluminescence analyzer provided in an embodiment of the present disclosure.

[0079] FIG4 shows a schematic structural diagram of a chemiluminescence analyzer provided in an embodiment of the present disclosure.

[0080] 5 to 15 are schematic block diagrams of photometers provided according to different embodiments of the present disclosure.

[0081] FIG16A to FIG16D are linear fitting curves of the luminescence values ​​of the chemiluminescent substrate solutions 1-1 to 1-4 of Example 1 in the alkaline phosphatase system.

[0082] FIG. 17A to FIG. 17D are linear fitting curves of the luminescence values ​​of the chemiluminescent substrate solutions 2-1 to 2-4 of Example 2 in the alkaline phosphatase system.

[0083] FIG. 18A to FIG. 18D are linear fitting curves of the luminescence values ​​of the chemiluminescent substrate solutions 3-1 to 3-4 of Example 3 in the alkaline phosphatase system.

[0084] FIG. 19A to FIG. 19D are linear fitting curves of the luminescence values ​​of the chemiluminescent substrate solutions 4-1 to 4-4 of Example 4 in the alkaline phosphatase system.

[0085] FIG. 20A to FIG. 20D are linear fitting curves of the luminescence values ​​of the chemiluminescent substrate solutions 5-1 to 5-4 of Example 5 in the alkaline phosphatase system. DETAILED DESCRIPTION

[0086] The following will clearly and completely describe the embodiments of the present disclosure in conjunction with the accompanying drawings. Obviously, the embodiments described are only part of the embodiments of the present disclosure, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present disclosure without making any creative efforts shall fall within the scope of protection of the present disclosure.

[0087] It should be noted that the terms "first\second\third" involved in the embodiments of the present disclosure are only used to distinguish similar objects and do not represent a specific order for the objects. It can be understood that "first\second\third" can be interchanged with a specific order or sequence when permitted.

[0088] Those skilled in the art will understand that unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs.

[0089] As mentioned in the background art, the HCG detection project has relatively high requirements for the upper limit of linear detection, but the existing technology cannot take into account the requirements of high linear detection upper limit and high detection sensitivity at the same time, and often requires dilution and re-testing to obtain accurate HCG concentration.

[0090] Based on this, the embodiments of the present disclosure propose a technical solution that can accurately and efficiently obtain HCG test results without increasing costs.

[0091] As shown in FIG1 , a first aspect of the present disclosure provides a chemiluminescence analyzer 100 , which includes a sample preparation device 110 , a substrate providing device 120 , a photometric device 130 , and a data processing device 140 .

[0092] The sample preparation device 110 is at least used to pre-dilute a blood sample to be tested for human chorionic gonadotropin using a diluent at a preset dilution ratio before the blood sample is tested for human chorionic gonadotropin. The pre-diluted blood sample is mixed with a reaction reagent in a reaction container so that human chorionic gonadotropin in the pre-diluted blood sample reacts with the reaction reagent to produce a test sample solution. The reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent carrying a labeled human chorionic gonadotropin antibody. In some embodiments, the capture reagent can be a magnetic bead reagent for capturing the substance to be tested, and the labeled detection reagent can be a detection reagent carrying an enzyme label, such as alkaline phosphatase.

[0093] The substrate providing device 120 is used to provide a luminescent substrate into a reaction container containing a sample solution to be tested, so that the marker in the sample solution to be tested reacts with the luminescent substrate to undergo a chemiluminescent reaction;

[0094] The photometric device 130 includes at least a photometer, which is used to detect the luminescent signal generated in the chemiluminescent reaction. The first linear detection range of the photometer is [A1, A2] photon counts / second, which represents the range of the number of photons output by the photometer. Within this first linear detection range, the intensity of the luminescent signal generated in the chemiluminescent reaction is linearly related to the photon count output by the photometer, wherein A1 is less than or equal to 2000 and A2 is greater than or equal to 10 8 .

[0095] The data processing device 140 is used to obtain the detection result of human chorionic gonadotropin in the blood sample to be tested according to the luminescence signal and a preset calibration model.

[0096] This design of the photometer results in a chemiluminescence analyzer with a wide linear detection range. Furthermore, by pre-diluting the sample, for example, by 2x, 5x, or 10x, a single test can yield a more accurate HCG test result, eliminating the need to pre-screen the sample and determine whether the sample should be tested with or without dilution. This can largely avoid unnecessary retesting and reduce the retest rate for samples with high HCG concentrations, thereby improving the overall efficiency of the chemiluminescence analyzer for HCG testing and reducing testing costs.

[0097] In particular, the chemiluminescence analyzer according to the present disclosure can accurately and linearly detect HCG as low as 0.5mIU / ml and as high as 200,000mIU / ml without the need for dilution and retesting. In other words, it is possible to accurately obtain HCG with a concentration between 0.5 and 200,000mIU / ml through a single test, meeting the main needs of clinical use of HCG to assist in determining whether pregnancy is present and monitoring pregnancy status, avoiding re-dilution and retesting when the sample HCG concentration exceeds the upper limit of detection, greatly improving the work efficiency of the laboratory and saving costs. As shown in Figures 2 and 3, for the HCG detection project, within the HCG concentration range of 0.5 to 200,000mIU / ml, the calibration model or calibration curve has good linearity in the low HCG concentration range, and also has good linearity in the entire HCG concentration range.

[0098] In the embodiment of the present disclosure, the linear detection range refers to the linear detection range of a single detection, that is, the linear detection range that can be achieved in one detection of one sample.

[0099] FIG4 is a schematic diagram of the structure of a chemiluminescence analyzer 100 provided in an embodiment of the present disclosure. As shown in FIG4 , the sample preparation device 110 includes a sample supply unit 111, a reagent supply unit 112, and a reaction incubation unit 113. The sample supply unit 111 is configured to draw a blood sample to be tested and supply it to a reaction vessel, and the reagent supply unit 112 is configured to capture a reagent and a detection reagent with a marker and supply them to the reaction vessel. The reaction incubation unit 113 is configured to provide a reaction and incubation area for the reaction vessel containing the blood sample to be tested and the reaction reagent, so that the blood sample to be tested and the reaction reagent in the reaction vessel form a sample solution to be tested.

[0100] The chemiluminescence analyzer 100 further includes a magnetic separation device 140 and a transport device 150. The magnetic separation device 140 is configured to perform a magnetic separation operation on a reaction container containing a sample solution to be tested. The substrate providing device 120 (not shown) is configured to provide a luminescent substrate to the reaction container that has undergone the magnetic separation operation. The transport device 150 is configured to transport the reaction container that has undergone the magnetic separation operation from the magnetic separation device 140 to the reaction incubation unit 113. After incubation, the reaction container is transported to the photometry device 130 for photometry.

[0101] In some embodiments, the chemiluminescence analyzer 100 further includes a sample handling system (not shown) for transporting test tubes containing blood samples to be tested to the sample aspiration position and for transporting test tubes after sample aspiration to a recovery area. Specifically, the sample handling system includes a sample loading area for storing test tubes containing blood samples to be tested inserted by the user, and a transport track for transporting test tubes from the sample loading area to the sample aspiration position.

[0102] In addition, the sample processing system also includes a scanning module for scanning sample barcodes to achieve sample identification and management.

[0103] In some embodiments, the sample supply unit 111 may include a sample needle and a drive mechanism for moving the sample needle between a sample aspiration position and a sample loading position. The sample needle aspirates a sample from a sample tube at the sample aspiration position and injects the aspirated sample into a reaction vessel at the sample loading position. The sample aspiration position may be, for example, the intersection of the sample needle's motion trajectory and the sample transport trajectory of the transport track.

[0104] In addition, the sample supply unit 111 also includes a cleaning mechanism for cleaning the sample needle. Furthermore, the sample supply unit 111 also includes a dilution position for placing a reaction container that has completed automatic dilution and pretreatment, so that the sample needle can aspirate the diluted sample or pretreated sample in the reaction container located in the dilution position to support the corresponding test mode.

[0105] In some embodiments, the chemiluminescence analyzer further includes a first manipulator 160 for transporting reaction vessels. The first manipulator is configured to move in three dimensions and to grip the reaction vessels. The first manipulator is configured to load a new reaction vessel into the sample loading position so that the sample needle can inject the sample. The first manipulator is also configured to discard an empty reaction cuvette. The first manipulator is further configured to transport the reaction vessel between the sample loading position, the reaction incubation section 113, and the dilution position.

[0106] In some embodiments, the reagent supply unit 112 includes a reagent tray 114 and a reagent needle 115. The reagent tray 114 is used to place reagent bottles containing capture reagents and detection reagents with markers and has the function of refrigerating the reagent bottles. The reagent tray 114 is configured to be rotatable so that reagent bottles that need to be aspirated can be transferred to the aspirating position. The reagent needle 115 is used to aspirate reagents from the reagent bottles located at the aspirating position and discharge the aspirated reagents into a reaction vessel located at the sample loading position.

[0107] Furthermore, the reagent tray has a mixing mechanism (not shown) for mixing the magnetic bead reagent. The mixing mechanism achieves mixing of the capture reagent by, for example, rotating a reagent bottle containing the capture reagent, such as the magnetic bead reagent.

[0108] Furthermore, the reagent supply unit 112 further includes a cleaning mechanism (not shown) for cleaning the reagent needle.

[0109] In some embodiments, the magnetic separation device 140 has a disc-shaped rotatable magnetic separation disc, a magnetic mechanism and a constant temperature control mechanism. The magnetic separation disc is used to drive the reaction containers therein to pass through the magnetic mechanism in sequence for magnetic separation operations, and the constant temperature control mechanism is used to keep the temperature of the magnetic separation disc within a preset range.

[0110] In some embodiments, the substrate providing device has a heating mechanism for preheating the luminescent substrate. The substrate providing device is used to inject the preheated luminescent substrate into a reaction vessel that has completed magnetic separation and is located at a substrate injection position, wherein the substrate injection position is located on the magnetic separation disk.

[0111] In some embodiments, the reaction incubation section 113 comprises a rotatable, disc-shaped reaction tray, with a light measuring device 130 disposed on the outer periphery of the reaction tray. After the reaction container is injected with the luminescent substrate, it is transported to the reaction tray by a transport device 150. The reaction tray carries the reaction container, which has been incubated for a certain period of time, into the light measuring device 130 for light measurement.

[0112] In some embodiments, the transfer device 150 is configured as a second robot arm, which is used to grip the reaction container and transfer the reaction container between the reaction disk and the magnetic separation disk. The second robot arm is configured to be able to rotate horizontally and to move in two dimensions.

[0113] In some embodiments, the chemiluminescence analyzer may further include a mixer 170 disposed between the reaction disk and the magnetic separation disk. The mixer is configured to mix the reaction vessels to which the reagents and samples have been added. The mixer may be configured, for example, as a vortex mixer for non-contact mixing, effectively preventing cross contamination.

[0114] The chemiluminescence analyzer may also include a hardware control system (not shown) and software systems running on the hardware board and PC, respectively, to control the coordinated operation of the various components of the chemiluminescence immunoassay. The chemiluminescence analyzer may also include a mechanical system (not shown) consisting of a frame and a housing to provide support, positioning, and protection for the various components.

[0115] In some embodiments, the data processing device 140 includes, but is not limited to, a central processing unit (CPU), a microcontroller unit (MCU), a field-programmable gate array (FPGA), a digital signal processing device (DSP), and other devices for interpreting computer instructions and processing data in computer software. For example, the data processing device is used to execute various computer applications stored in a computer-readable storage medium, thereby causing the chemiluminescence analyzer 100 to execute the corresponding detection process and analyze the luminescence signal detected by the photometric device 130 in real time.

[0116] A specific detection process of the chemiluminescence analyzer is as follows: the sample supply unit 111 draws the blood sample to be tested and adds it to the reaction container; the reagent supply unit 112 draws the capture reagent and detection reagent and adds them to the reaction container with the added sample to mix with the sample; the reaction container is then placed in the reaction incubation unit 113 for reaction, incubation and magnetic separation and cleaning; then the substrate supply device adds the luminescent substrate to the reaction cup that has completed the reaction, incubation and magnetic separation and cleaning, and incubates for a period of time; finally, the photometric device 130 detects the photons emitted by the analyte in the sample under the action of the luminescent substrate, so that the concentration level of the analyte can be calculated based on the measured number of photons.

[0117] In some embodiments, the preset dilution ratio includes 2 to 20 times. This can improve the applicability of the analyzer for samples of different concentrations. In other words, for samples with a wide range of HCG concentrations, more accurate HCG detection results can be obtained, thereby improving the ability of the chemiluminescence analyzer to detect HCG.

[0118] In some embodiments, a calibration model is generated by using a pre-diluted calibrator with a chemiluminescence analyzer. Calibration using a pre-diluted calibrator involves directly tracing the pre-diluted calibrator back to the standard, and establishing a gradient concentration response curve for the pre-diluted calibrator. This eliminates systematic errors caused by random bias in the dilution process and matrix effects, thereby improving measurement accuracy.

[0119] In some embodiments, the pre-dilution factor of the blood sample to be tested is equal to the pre-dilution factor of the calibrator. In this way, the matching degree between the calibration model and the actual test results can be improved, thereby more accurately obtaining the actual HCG content in the blood sample to be tested.

[0120] In some embodiments, the diluent, capture reagent and detection reagent are placed in a common kit, which includes a carrier and a first cavity, a second cavity and a third cavity opened on the carrier, the first cavity is used to hold the diluent, the second cavity is used to hold the capture reagent, and the third cavity is used to hold the detection reagent.

[0121] This results in the following benefits:

[0122] a) Automatic operation of HCG dilution on the machine;

[0123] b) The manufacturer provides diluent, which can ensure the accuracy of HCG dilution measurement and avoid poor test results caused by customers preparing diluent on their own;

[0124] c) Save reagent storage space and improve reagent space utilization efficiency;

[0125] d) It saves testing costs and provides great convenience to customers as they do not need to take inventory or purchase diluents separately.

[0126] In some embodiments, the calibration model or calibration curve is obtained using multi-point calibration.

[0127] In the embodiment of the present disclosure, the multi-point calibration method can be: using a series of samples (greater than or equal to 5 samples) pre-assigned by the supplier of the chemiluminescence analyzer, measuring the luminescence signal values ​​of this series of samples on the chemiluminescence analyzer, establishing a curve of sample concentration and luminescence signal, and obtaining a fitting equation, i.e., a calibration model, by performing 4PLC fitting regression on the curve.

[0128] It can be understood that the luminescence signal value of the blood sample to be tested measured on the chemiluminescence analyzer is substituted into the calibration model to calculate the concentration of the substance to be tested in the blood sample to be tested.

[0129] In some embodiments, the calibration model is obtained by the following formula

[0130] Among them, RLU is the reactivity, that is, the measured luminescence signal value, C is the concentration of the substance being measured, P1 to P4 are four fitting parameters, among which P1 represents the luminescence signal corresponding to zero concentration (asymptote), P4 represents the luminescence signal corresponding to the maximum concentration (asymptote), P2 represents the inflection point where the slope direction of the curve changes, and P3 reflects the slope at P2 (not the true slope).

[0131] It can be understood here that the 4PLC calibration model uses at least 5 (number of fitting parameters + 1) points for 4PLC regression, and obtains the optimal solution through a numerical iteration process to obtain four fitting parameters P1, P2, P3, and P4.

[0132] In some embodiments of the present disclosure, the calibration model is recalibrated when the reagent batch is changed, or the calibration model is recalibrated at fixed intervals of days, or the calibration model is recalibrated when the instrument quality control is out of control.

[0133] In some embodiments, the ratio of A2 to A1 (A2 / A1) is greater than or equal to 30,000.

[0134] In some embodiments, the second linear detection range of the luminescent substrate is [B1, B2] photon counts / second, wherein the second linear detection range represents the range of the intensity of the luminescent signal generated in the chemiluminescent reaction. Within the second linear detection range, the number of markers participating in the chemiluminescent reaction is linearly related to the intensity of the luminescent signal generated in the chemiluminescent reaction, wherein B1 is less than or equal to 3000 and B2 is greater than or equal to 10 8 , and / or the ratio of B2 to B1 B2 / B1 is greater than or equal to 30000.

[0135] Here, by designing the luminometer and the luminescent substrate in this way, a chemiluminescent analyzer with a wide linear detection range can be obtained, in particular, a chemiluminescent analyzer with a linear detection range of [3000, 10 8 ] photon counts / second in the linear detection range of the chemiluminescence analyzer.

[0136] In some embodiments, the lower limit B1 of the second linear detection range is less than or equal to 2000. This can further improve the sensitivity of the chemiluminescence analyzer, thereby further broadening the linear detection range of the chemiluminescence analyzer for single detection.

[0137] In some embodiments, the chemiluminescence analyzer 100 can be designed such that when the concentration of human chorionic gonadotropin in the blood sample to be tested is greater than or equal to 200,000 mIU / ml, the intensity of the luminescent signal generated in the chemiluminescent reaction is less than or equal to the smaller value of A2 and B2. This allows for more accurate detection of higher concentrations of HCG.

[0138] For example, the chemiluminescence analyzer 100 can be designed such that, when the concentration of human chorionic gonadotropin in the blood sample to be tested is 200,000 mIU / ml, the intensity of the luminescent signal generated in the chemiluminescent reaction is less than or equal to the smaller value of A2 and B2. This enables accurate detection of HCG concentrations up to 200,000 mIU / ml.

[0139] In some embodiments, the chemiluminescence analyzer 100 can be designed such that when the concentration of human chorionic gonadotropin in the blood sample to be tested is greater than or equal to 0.5 mIU / ml, the intensity C1 of the luminescent signal generated in the chemiluminescent reaction is greater than or equal to the larger value of A1 and B1. This allows for more accurate detection of HCG at lower concentrations.

[0140] For example, the chemiluminescence analyzer 100 can be designed such that, when the concentration of human chorionic gonadotropin in the blood sample to be tested is 0.5 mIU / ml, the intensity C1 of the luminescent signal generated in the chemiluminescent reaction is greater than or equal to the larger value of A1 and B1. This enables accurate detection of HCG concentrations as low as 0.5 mIU / ml.

[0141] Furthermore, the chemiluminescence analyzer 100 can be designed such that, when the concentration of human chorionic gonadotropin in the blood sample to be tested is greater than 0.5 mIU / ml and less than or equal to 5 mIU / ml, preferably less than or equal to 2 mIU / ml, the intensity of the luminescent signal generated in the chemiluminescence reaction is C2, wherein the ratio of C2 to C1, C2 / C1, is greater than 2.

[0142] In some embodiments, the upper limit B2 of the second linear detection range is greater than the upper limit A2 of the first linear detection range.

[0143] Next, some examples of luminescent substrates used in the chemiluminescent analyzer of the present disclosure are described, but the present disclosure is not limited thereto.

[0144] In some embodiments, the luminescent substrate is designed such that the background of the luminescent substrate is less than 3000, thereby further improving the sensitivity of the chemiluminescent analyzer.

[0145] Alternatively or additionally, the luminescent substrate is designed such that the signal-to-noise ratio of the luminescent substrate is greater than 40000. This can further improve the sensitivity of the chemiluminescent analyzer.

[0146] In some embodiments, the luminescent substrate comprises a chemiluminescent substrate and a chemiluminescence enhancer, the chemiluminescence enhancer comprises a fluorescent agent and a surfactant, and the fluorescent agent comprises one or more carboxyfluoresceins. Using such a luminescent substrate can achieve high sensitivity of a chemiluminescence analyzer.

[0147] As some implementations, the luminescent substrate is selected from dioxetane compounds, preferably one or more selected from AMPPD, CSPD, CDP-STAR, and more preferably CDP-STAR.

[0148] As some implementations, the luminescent substrate is selected from chlorinated derivatives of AMPPD, fluorescein or its carboxyl-substituted derivatives, and water-soluble polymeric quaternary ammonium salt-type cationic surfactants.

[0149] The chlorinated derivative of AMPPD may be, for example, ADP-STAR or CDP-STAR. Alternatively, the fluorescein or its carboxyl-substituted derivative may be, for example, fluorescein or 5(6)-carboxyfluorescein. Alternatively, the water-soluble polymeric quaternary ammonium salt cationic surfactant may be, for example, polyvinylbenzyltrimethylammonium chloride.

[0150] In some embodiments, the substrate providing device is used to provide a chemiluminescent substrate liquid containing a luminescent substrate or a chemiluminescent substrate, wherein the chemiluminescent substrate liquid includes a chemiluminescent substrate, fluorescein and a water-soluble polymer quaternary ammonium salt, wherein the chemiluminescent substrate is selected from chlorinated dioxetane compounds having a spiro-adamantane substituent.

[0151] Chemiluminescent substrates are compounds that participate in energy transfer in a chemiluminescent reaction and ultimately release energy in the form of emitted photons. They are also called chemiluminescent agents or luminescent substrates. Dioxetanes are enzymatic glow-type chemiluminescent substrates and are ultrasensitive substrates for alkaline phosphatase (AP). In a suitable buffer, the signal emitted by the decomposition of dioxetanes, catalyzed by alkaline phosphatase, can last for over 20 hours, making them ideal chemiluminescent substances.

[0152] The present inventors have discovered that chlorinated dioxetanes having spiro-adamantane substituents, when combined with specific chemiluminescence enhancers, such as fluorescein or its carboxylic acid derivatives, in the presence of a water-soluble polymeric quaternary ammonium salt cationic surfactant, can produce a chemiluminescent substrate solution with a photon number per second ratio of the upper detection limit to the lower detection limit of more than 30,000, or even more than 50,000. This allows for convenient application in samples such as HCG with a concentration range of up to 10 5 ~10 6 Order of magnitude immunoassay.

[0153] The "dioxetanes having spiro-adamantane substituents" mentioned herein refer to a class of compounds based on the following structure:

[0154] For example, the following compounds are included, but not limited to:

[0155] AMPPD-(3-(2'-spiroadamatane)-4-methoxy-4-(3"-phosphoryloxy)-phenyl-1,2-dioxetane,CAS=122341-56-4),

[0156] CSPD-(3-(2'-(spiro-5-chloroadamantane))-4-methoxy-4-(3"-phosphoryloxy)-phenyl-1,2-dioxetane,CAS=142456-88-0),

[0157] ADP-STAR-(3-(2'-spiroadamatane)-4-methoxy-4-(3"-phosphoryloxy-4"-chloro)-phenyl-1,2-dioxetane, CAS=189942-84-5),

[0158] CDP-STA-(3-(2'-(spiro-5-chloroadamantane))-4-methoxy-4-(3"-phosphoryloxy-4"-chloro)-phenyl-1,2-dioxetane, CAS=160081-62-9),

[0159] TFE-AMPPD-(3-(2'-spiroadamatane)-4-trifluoroethoxy-4-(3"-phosphoryloxy)-phenyl-1,2-dioxetane).

[0160] The "chlorinated dioxetane compound having a spiro-adamantane substituent" mentioned herein refers to a compound containing at least one chlorine atom in the structure of AMPPD.

[0161] In some embodiments, the chlorinated dioxetane compound having a spiro-adamantane substituent is one of ADP-STAR and CDP-STAR.

[0162] In a specific embodiment, in the chemiluminescent substrate solution, the chemiluminescent substrate may be present in the form of a salt of the above-mentioned compound. According to some embodiments, the salt may be an alkali metal salt, such as a sodium salt.

[0163] These dioxetane compounds with chlorinated groups, especially ADP-STAR and CDP-STAR, when combined with fluorescein or its carboxyl derivatives and water-soluble polymer quaternary ammonium salts in an alkaline phosphatase catalytic system, unexpectedly provide sufficiently low sensitivity while maintaining linearity in a higher luminescence value range.

[0164] Fluorescent agents and surfactants can enhance chemiluminescence efficiency. Surfactants form micelles in solution, protecting the chemiluminescent substrate and reducing its quenching reaction in aqueous solution. Fluorescent agents act as photon receptors, receiving photon energy through energy transfer and being excited to generate light signals, thereby enhancing luminescence efficiency.

[0165] The fluorescent agent in the chemiluminescent substrate solution disclosed herein is at least one of fluorescein and carboxyl-substituted fluorescein.

[0166] The carboxyl-substituted fluorescein includes compounds selected from the group consisting of:

[0167] Specifically, the carboxyfluorescein represented by general formula I can be 5-carboxyfluorescein, 6-carboxyfluorescein, or 5(6)-carboxyfluorescein.

[0168] The 5-carboxyfluorescein is

[0169] The 6-carboxyfluorescein is

[0170] The 5(6)-carboxyfluorescein is a mixture of the 5-carboxyfluorescein and the 6-carboxyfluorescein in any proportion.

[0171] In the chemiluminescent substrate solution, the fluorescent agent is fluorescein. In other embodiments, the fluorescent agent can be any carboxyl-substituted fluorescein, in particular one of 5-carboxyfluorescein, 6-carboxyfluorescein and 5(6)-carboxyfluorescein.

[0172] Fluorescein and its carboxyl derivatives can improve the chemiluminescence efficiency of chemiluminescent systems. In addition to effectively improving the chemiluminescence efficiency of chemiluminescent systems, these carboxyl derivatives can also reduce the time required to reach the plateau phase and improve sensitivity.

[0173] In the described chemiluminescent substrate liquid, the surfactant is a water-soluble polymer quaternary ammonium salt cationic surfactant. According to one embodiment, the water-soluble polymer quaternary ammonium salt is selected from a quaternary ammonium salt containing a polyvinyl group. This type of quaternary ammonium salt, for example, is a polyvinyl benzyl-trialkyl ammonium halide. The three alkyl groups substituted on the N atom can be the same or different, for example, can independently be selected from methyl, ethyl, propyl, butyl, amyl, benzyl, etc. In a specific embodiment, the water-soluble polymer quaternary ammonium salt is selected from at least one of polyvinyl benzyl-trimethyl ammonium chloride, polyvinyl benzyl-benzyldimethyl ammonium chloride and polyvinyl benzyl-tributylammonium chloride.

[0174] Water-soluble high-molecular-weight quaternary ammonium salts, such as cationic surfactants, can also form micelles in aqueous solutions. The chemiluminescent substrate is located within the hydrophobic micelles, while the fluorescent agent is located in the hydrophilic portion of the micelles. This facilitates the transfer of energy from the chemiluminescent substrate to the fluorescent agent, thereby enhancing the light signal. Compared with small-molecule quaternary ammonium salts, high-molecular-weight quaternary ammonium salts have longer alkane chains, which can more effectively prevent quenching of the chemiluminescent substrate in aqueous solutions. Furthermore, high-molecular-weight quaternary ammonium salts can also suppress background signals, thereby increasing detection signals. However, under the same conditions, small-molecule quaternary ammonium salts have high background signal values ​​and low detection signals, making it impossible to achieve a wide linear detection range.

[0175] In one embodiment, the chemiluminescent substrate solution includes ADP-STAR or CDP-STAR as a chemiluminescent substrate, fluorescein and a water-soluble polymer quaternary ammonium salt.

[0176] In another embodiment, the chemiluminescent substrate solution includes ADP-STAR, fluorescein or carboxyl-substituted fluorescein as a fluorescent agent, and a water-soluble polymer quaternary ammonium salt.

[0177] The specific combination of the above chemiluminescent substrate, fluorescent agent, and surfactant provides a chemiluminescent substrate solution with a wide linear luminescence value. The chemiluminescent substrate solution can obtain a linear detection range with a lower limit B1 of less than 3,000 photon counts / second and an upper limit B2 of more than 100,000,000 (100M) photon counts / second in an alkaline phosphatase system. Exemplarily, the chemiluminescent substrate solution can obtain a linear detection range with a lower limit B1 of 1,000 to 3,000 photon counts / second and an upper limit B2 of 100,000,000 (100M) to 200,000,000 (200M) photon counts / second in an alkaline phosphatase system. As shown in the following examples, the chemiluminescent substrate solution can achieve a background luminescence value of 1,000 to 2,000 photon counts / second and a linear detection range of an upper limit B2 of 120,000,000 (120M) to 200,000,000 (200M) photon counts / second in an alkaline phosphatase system. It should be understood that the above lower and upper limits are merely exemplary, and lower photon counts / second lower limits or higher photon counts / second upper limits can be achieved by adjusting the detection equipment or signal processing methods.

[0178] In some embodiments, the ratio of the number of photons per second of the upper detection limit B2 of the chemiluminescent substrate solution to the lower detection limit B1 is greater than 30,000, for example, greater than 50,000, or even greater than 60,000. In some embodiments, the ratio of the number of photons per second of the upper detection limit B2 of the chemiluminescent substrate solution to the lower detection limit B1 is less than 1 million, for example, less than 800,000, less than 600,000, less than 500,000, or even less than 400,000. Exemplarily, the ratio of the number of photons per second of the upper detection limit B2 of the chemiluminescent substrate solution to the lower detection limit B1 is 30,000 to 1 million, 30,000 to 800,000, 30,000 to 500,000, 50,000 to 1 million, 50,000 to 800,000, 50,000 to 500,000, etc.

[0179] In some embodiments, the chemiluminescent substrate solution comprises 50-500 mg / L of chemiluminescent substrate. Preferably, the chemiluminescent substrate solution comprises 150-250 mg / L of chemiluminescent substrate, and more preferably, comprises 200 mg / L of chemiluminescent substrate. Exemplary, the chemiluminescent substrate solution comprises 150 mg / L, 160 mg / L, 170 mg / L, 180 mg / L, 190 mg / L, 200 mg / L, 210 mg / L, 220 mg / L, 230 mg / L, 240 mg / L, or 250 mg / L of chemiluminescent substrate.

[0180] In some embodiments, the chemiluminescent substrate solution comprises 30 to 500 mg / L of a fluorescent agent. Preferably, the chemiluminescent substrate solution comprises 150 to 250 mg / L of a fluorescent agent, more preferably, 200 mg / L of a fluorescent agent. Exemplary, the chemiluminescent substrate solution comprises 150 mg / L, 160 mg / L, 170 mg / L, 180 mg / L, 190 mg / L, 200 mg / L, 210 mg / L, 220 mg / L, 230 mg / L, 240 mg / L, or 250 mg / L of a fluorescent agent.

[0181] In some embodiments, the chemiluminescent substrate solution comprises 1-10 g / L of a water-soluble polymeric quaternary ammonium salt. Preferably, the chemiluminescent substrate solution comprises 3-7 g / L of a water-soluble polymeric quaternary ammonium salt, and more preferably, comprises 5 g / L of a water-soluble polymeric quaternary ammonium salt. Exemplarily, the chemiluminescent substrate solution comprises 3 g / L, 4 g / L, 5 g / L, 6 g / L, or 7 g / L of a water-soluble polymeric quaternary ammonium salt.

[0182] In one embodiment, the chemiluminescent substrate solution includes 50-500 mg / L, preferably 150-250 mg / L of ADP-STAR or CDP-STAR, 30-500 mg / L, preferably 150-250 mg / L of fluorescein, and 1-10 g / L, preferably 3-7 g / L of a water-soluble polymer quaternary ammonium salt.

[0183] In another specific embodiment, the chemiluminescent substrate solution includes 50-500 mg / L, preferably 150-250 mg / L of ADP-STAR, 30-500 mg / L, preferably 150-250 mg / L of fluorescein or carboxyl-substituted fluorescein and 1-10 g / L, preferably 3-7 g / L of water-soluble polymer quaternary ammonium salt.

[0184] The chemiluminescent substrate solution also contains additives such as a buffer and a preservative.

[0185] The present disclosure does not particularly limit the type of buffer, and conventionally applicable buffers can be used for the chemiluminescent substrate solution of the present disclosure. In some embodiments, the buffer can be selected from Tris buffer, AMP buffer (AMP is 2-amino-2-methylpropanol (2-amino-2-methylpropanol)), AMPD buffer (AMPD is 2-amino-2-methyl-1,3-propanediol (2-amino-2-methyl-1,3-propanediol)), DEA buffer (DEA is diethanolamine (diethanolamine)), CHES buffer (CHES is 2-(N-cyclohexylamino)ethanesulfonic acid (2-(N-cyclohexylamino)ethanesulfonic acid)), Mopso buffer, imidazole buffer, phosphate buffer, carbonate buffer, malic acid buffer, glycine buffer, but is not limited thereto. Exemplarily, the buffer can be selected from at least one of Tris-HCl, AMP-HCl, AMPD-HCl, DEA-HCl, CHES-HCl, boric acid-NaOH, and glycine-NaOH buffer systems. Preferably, the buffer is selected from the AMP-HCl buffer system. The AMP-HCl buffer system can play a good buffering role in the pH range of 9.0 to 10.0, and studies have found that it is beneficial to the stability of the chemiluminescent substrate molecules.

[0186] According to some embodiments, the pH of the buffer is 7.1 to 10.6, preferably about 9.0 to about 10.0, and particularly preferably about 9.5.

[0187] The present disclosure has no particular limitation on the amount of the buffer used, which can be determined according to the selected buffer system.

[0188] The present disclosure does not particularly limit the type of preservative; any preservative conventionally used in detection reagents can be used in the present disclosure. Examples include, but are not limited to, sodium azide, Proclin series preservatives, potassium sorbate, sodium benzoate, BND, erythromycin, and gentamicin. Preferred preservatives include sodium azide and Proclin series preservatives.

[0189] The present disclosure has no particular limitation on the amount of the buffer, which can be determined according to the type of preservative selected.

[0190] In some embodiments, the chemiluminescent substrate solution may further include magnesium chloride, for example, 0.1 to 2 mM magnesium chloride.

[0191] According to a specific embodiment, the chemiluminescent substrate solution includes 50-500 mg / L, preferably 150-250 mg / L of ADP-STAR or CDP-STAR; 30-500 mg / L, preferably 150-250 mg / L of fluorescein; 1-10 g / L, preferably 3-7 g / L of a water-soluble polymer quaternary ammonium salt; an appropriate amount (such as 30-500 mM) of AMP-HCl buffer; an appropriate amount (such as 0.5-2 g / L) of sodium azide, and the pH is about 9.0 to about 10.0, such as about 9.5.

[0192] In another specific embodiment, the chemiluminescent substrate solution includes 50-500 mg / L, preferably 150-250 mg / L of ADP-STAR; 30-500 mg / L, preferably 150-250 mg / L of fluorescein or carboxyl-substituted fluorescein; 1-10 g / L, preferably 3-7 g / L of a water-soluble high molecular weight quaternary ammonium salt; an appropriate amount (such as 30-500 mM) of AMP-HCl buffer; an appropriate amount (such as 0.5-2 g / L) of sodium azide, and the pH is about 9.0 to about 10.0, such as about 9.5.

[0193] According to some embodiments, the chemiluminescent substrate solution disclosed herein has low background luminescence value and high luminescence efficiency, a wide linear detection range, is suitable for alkaline phosphatase enzymatic luminescence system, and is particularly suitable for chemiluminescent immunoassay.

[0194] According to some specific embodiments, the chemiluminescent substrate solution provided by the present disclosure reacts with alkaline phosphatase in a concentration range of 0.03 to 400 ng / mL, and the luminescence value shows good linearity (R2 ≥ 0.99). As shown in the examples, the background value of the chemiluminescent substrate solution can be as low as 3,000 photon counts / second or less, and can detect ultra-low concentrations of alkaline phosphatase as low as 10-19 mol / L, thereby improving the detection sensitivity; at the same time, the detection value of the chemiluminescent substrate solution can be as high as 100M or more photon counts / second, for example, 100M to 200M photon counts / second, 120M to 200M photon counts / second, showing a significantly wider linearity. Without additional dilution of the sample, the above-mentioned chemiluminescent substrate solution can meet the detection of samples with high analyte concentrations.

[0195] The chemiluminescent substrate solution disclosed herein is particularly suitable for detecting human chorionic gonadotropin (HCG) in a sample to be tested.

[0196] HCG is a glycoprotein primarily secreted by placental trophoblast cells. Its concentration in non-pregnant individuals is very low, typically less than 3 mIU / ml. During pregnancy, HCG concentrations can reach as high as approximately 2,000,000 mIU / ml. Clinically, HCG test results are used to assist in determining pregnancy. Pregnancy also plays an important role in HCG testing. As previously mentioned, the chemiluminescent substrate solution disclosed herein exhibits broad linear luminescence characteristics, making it suitable for samples with wide variations in analyte concentrations, such as HCG.

[0197] According to some embodiments, the concentration of HCG in the sample ranges from 1 to 200,000 mIU / mL.

[0198] In some embodiments, HCG is detected using a double antibody sandwich method. Depending on the specific embodiment, the detection uses superparamagnetic particles coated with HCG antibodies as the capture reagent, and alkaline phosphatase-labeled HCG antibodies as the marker. In some embodiments, the HCG antibodies are monoclonal antibodies.

[0199] The chemiluminescent substrate solution combined with the double antibody sandwich method disclosed in the present invention has high sensitivity for HCG detection and a wide linear detection range.

[0200] The present disclosure does not particularly limit the specific form and dosage of the chemiluminescent substrate solution used in the immunoassay of HCG, such as HCG antibodies, specific capture reagents, alkaline phosphatase markers, etc. Any suitable reagent can be used.

[0201] Unless otherwise specified, the "sample" mentioned herein refers to a biological sample, which may be from a mammal, preferably a blood sample from a human, and more preferably a serum sample.

[0202] In some embodiments, the chemiluminescent substrate solution is used to perform a chemiluminescent detection method in a chemiluminescent analyzer according to an embodiment of the present disclosure. The chemiluminescent detection method includes the following steps:

[0203] Mixing the sample to be tested with a detection reagent, wherein the detection reagent includes a capture reagent capable of binding to HCG and an alkaline phosphatase marker, thereby obtaining an alkaline phosphatase-labeled immune complex;

[0204] Mixing the alkaline phosphatase-labeled immune complex with the chemiluminescent substrate solution in any of the above embodiments to obtain a mixed solution;

[0205] The optical signal of the mixed solution is measured, and an analysis result of the sample to be tested is obtained based on the optical signal. Next, some embodiments of the photometer of the chemiluminescence analyzer of the present disclosure are described, but the present disclosure is not limited thereto.

[0206] In some embodiments, as shown in FIG5 , the photometer includes a receiving component 10 and a processing component 20 electrically connected to each other.

[0207] The receiving component 10 is configured to receive the light signal generated in the chemiluminescent reaction and convert the light signal into a corresponding electrical signal. The receiving component 10 can be configured as a photomultiplier tube that converts a weak light signal into an electrical signal.

[0208] The processing component 20 is configured to be electrically connected to the receiving component 10 to receive an electrical signal from the receiving component 10 and process the received electrical signal to obtain a photon count. The processing component 20 includes a first photon counting module 21 and a second photon counting module 22. The first photon counting module 21 is configured to process the electrical signal using a first photon counting method to obtain a first photon count result, while the second photon counting module 22 is configured to process the electrical signal using a second photon counting method different from the first photon counting method to obtain a second photon count result. The processing component 20 is further configured to output a final photon count result for the sample under test based on the first and second photon count results and use this as the photon count output by the photometer. This final photon count result, i.e., the photon count output by the photometer, can then be used to calculate the content of the substance under test in the sample under test.

[0209] In the embodiment of the present disclosure, two photon counting modules are used to count photons simultaneously, wherein the first photon counting module uses a first counting method for a weak light segment to obtain a first photon counting result, and the second photon counting module uses a second counting method for a strong light segment to obtain a second photon counting result. Finally, the first photon counting result and the second photon counting result are integrated and the final counting result is output. This can greatly expand the linear detection range of the photometer. In particular, it can achieve the linear detection range of the photometer to at least [2000, 100 million (10 8 )】Photon counts / second, even extended to 【2000, 200 million (10 8 )】Photon counts / second.

[0210] In the disclosed embodiments, a weak light segment can be understood as an optical signal having a photon count per second no greater than a predetermined threshold, while a strong light segment can be understood as an optical signal having a photon count per second greater than the predetermined threshold. For example, the predetermined threshold can be selected from any value between 20 million and 30 million photon counts per second, such as 20 million or 30 million photon counts per second. For example, the first counting method is a photon counting method for optical signals having a photon count per second no greater than 30 million photon counts per second, while the second counting method is a photon counting method for optical signals having a photon count per second greater than 30 million photon counts per second.

[0211] In some embodiments, the first counting method can be a pulse recognition method that calculates the photon count by identifying the electrical pulses caused by photons entering the receiving component, while the second counting method does not calculate the photon count by identifying the electrical pulses caused by photons entering the receiving component, but estimates the photon count by processing the electrical signal into parameters that can characterize the number of photons in the optical signal (i.e., the number of electrical pulses caused by photons entering the receiving component).

[0212] Here, those skilled in the art can understand that in the embodiment of the present disclosure, the pulse recognition method can be understood as a method of calculating photon counts by identifying pulses in electrical signals, that is, when the peak value of the pulse identified from the electrical signal is greater than a threshold, it is considered that a photon is identified.

[0213] In some embodiments, the first photon counting module 21 can be configured to detect the number of pulses of the electrical signal using a pulse recognition method to obtain a first photon counting result, and the second photon counting module 22 can be configured to process the electrical signal to obtain parameters characterizing the number of photons in the optical signal, and obtain, for example, calculate, the second photon counting result based on the parameters characterizing the number of photons in the optical signal and a preset calibration function, wherein the calibration function represents the mapping relationship between the parameters characterizing the number of photons in the optical signal and the photon counting result.

[0214] In some embodiments, after obtaining the first photon counting result and the second photon counting result, the processing component 20 may be further configured to:

[0215] When the first photon counting result is lower than a first threshold, taking the first photon counting result as a final photon counting result, that is, a photon count output by the photometer; and

[0216] When the first photon counting result is not lower than the first threshold, the second photon counting result is used as the final photon counting result, that is, the photon counting output by the photometer.

[0217] In some embodiments, as shown in FIG6 , the first photon counting module 21 may include a level discrimination circuit 211, a shaping and frequency division circuit 212, and a first counting circuit 213 electrically connected to each other. The level discrimination circuit 211 is configured to convert an electrical signal into a square wave signal, the shaping and frequency division circuit 212 is configured to perform frequency division and shaping on the square wave signal, and the first counting circuit 213 is configured to count the signal output by the shaping and frequency division circuit 212, i.e., identify the number of pulses, to obtain a first photon count result. The first photon counting module 21 according to this example can achieve a linear detection range of 2000 to 30 million photon counts / second.

[0218] In other alternative embodiments, as shown in FIG7 , the first photon counting module 21 may include at least one first AD conversion circuit 214 and a first counting circuit 213 electrically connected to each other. The first AD conversion circuit 214 is configured to collect electrical signals at a sampling frequency greater than 1 GHz / s and convert the collected electrical signals into digital signals, which are output to the first counting circuit 213. The first counting circuit 213 is configured to analyze the received digital signals to identify and receive the number of pulses, thereby obtaining a first photon counting result. Thus, by sampling at a high speed of 1 GHz / s, the first counting circuit 213 can more accurately identify the number of pulses in the electrical signal, thereby obtaining a more accurate first photon counting result. Compared to the embodiment shown in FIG6 , the first photon counting module 21 according to this embodiment can achieve a linear detection range of 2000 to 80 million photon counts / second.

[0219] In some variations of the embodiment shown in FIG7 , the first AD conversion circuit may be configured to acquire electrical signals at a sampling frequency of not less than 2 GHz / s, thereby further broadening the linear detection range of the first photon counting module 21 .

[0220] In some variations of the embodiment shown in FIG. 7 , the first photon counting module 21 may include at least two, and particularly at least four, first AD conversion circuits 214. The sum of the sampling frequencies of the first AD conversion circuits is no less than 1 GHz / s, and the first AD conversion circuits acquire electrical signals asynchronously at predetermined intervals. By using multiple first AD conversion circuits to achieve a sampling frequency of no less than 1 GHz / s, the performance requirements for a single AD conversion circuit can be reduced, thereby reducing costs.

[0221] Preferably, the first AD conversion circuits 214 have the same sampling frequency, and each first AD conversion circuit collects the electrical signal in an asynchronous manner at equal time intervals.

[0222] Accordingly, the first counting circuit 213 may be configured to rearrange and combine the data collected by the plurality of first AD conversion circuits 214 , perform filtering processing on the data, and then perform pulse recognition on the filtered signal to obtain a first photon counting result.

[0223] Furthermore, the first counting circuit 213 may be configured to correct the first photon counting result using a Poisson distribution compensation algorithm, thereby obtaining a more accurate photon counting result.

[0224] In some embodiments, the first counting circuit 213 may include an FPGA chip and its peripheral circuits.

[0225] In some embodiments, the second photon counting module 22 can be configured to process the electrical signal, for example, by integration, to obtain a DC component signal. The parameter characterizing the number of photons in the optical signal includes a parameter related to the DC component signal, such as the DC component signal itself per unit time. The second photon counting module 22 is further configured to obtain a second photon counting result based on the parameter related to the DC component signal and a preset calibration function.

[0226] Alternatively or additionally, the second photon counting module 22 is configured to integrate the electrical signal over a predetermined time period to obtain an integration result, such as a DC component signal. The parameter characterizing the number of photons in the optical signal includes a parameter related to the integration result, such as an integration result per unit time. The second photon counting module 22 is further configured to obtain a second photon counting result based on the parameter related to the integration result and a preset calibration function.

[0227] In a specific example, as shown in Figures 8 and 9, the second photon counting module 22 includes an integration circuit 221 and a second counting circuit 222 electrically connected to each other. The integration circuit 221 is configured to integrate the electrical signal over a predetermined time period to obtain a DC component signal, wherein the parameter representing the number of photons in the optical signal includes a parameter related to the DC component signal. The second counting circuit 222 is configured to obtain a second photon counting result based on the parameter related to the DC component signal and a preset calibration function.

[0228] Furthermore, as shown in FIG10 , an AD conversion circuit 224 is provided between the integration circuit 221 and the second counting circuit 222. The AD conversion circuit 224 is configured to convert the DC component signal output by the integration circuit 221 into a digital signal. The sampling frequency of the AD conversion circuit 224 can be set to less than 1 MHz / s, for example, several hundred kHz / s.

[0229] In some embodiments, the second counting circuit 222 may include an FPGA chip and its peripheral circuits.

[0230] Preferably, as shown in Figures 11 and 12, the first counting circuit 213 and the second counting circuit 222 are the same counting circuit, which may include, for example, an FPGA chip. That is, the counting functions of the first photon counting module 21 and the second photon counting module 22 are integrated into the same FPGA chip. Specifically, the FPGA chip is configured to count the signal output by the shaping and frequency dividing circuit 212 to obtain a first photon counting result (Figure 11) or analyze the digital signal collected by the first A / D conversion circuit 214 to obtain a first photon counting result (Figure 12). Furthermore, the FPGA chip is configured to obtain a second photon counting result based on a parameter representing the number of photons in the optical signal and a preset calibration function.

[0231] In another alternative example, as shown in Figures 13 and 14, the second photon counting module includes a second AD conversion circuit 223 and a second counting circuit 222 electrically connected to each other. The second AD conversion circuit is configured to convert the electrical signal output by the receiving component 10 into a digital signal, and the second counting circuit is configured to process the digital signal to obtain a parameter representing the number of photons in the optical signal, and obtain a second photon counting result based on the parameter representing the number of photons in the optical signal and a preset calibration function.

[0232] Here, a second AD conversion circuit 223 with a sampling frequency less than 10 MHz / s may be used. In some embodiments, the sampling frequency of the second AD conversion circuit 223 may be in the range of 1 MHz / s to 10 MHz / s, for example, 1, 2, or 3 MHz / s.

[0233] Preferably, as shown in Figure 15, the first counting circuit 213 and the second counting circuit 222 are the same counting circuit, which includes, for example, an FPGA chip. In other words, the counting functions of the first photon counting module 21 and the second photon counting module 22 are integrated into the same chip.

[0234] Also preferably, as shown in FIG. 15 , the first AD conversion circuit 214 is used as the second AD conversion circuit 223 , that is, the first photon counting module 21 and the second photon counting module 22 use the same AD conversion circuit.

[0235] Furthermore, the second counting circuit 222 may be configured to process the digital signal to obtain a DC component signal, and the parameter characterizing the number of photons in the optical signal includes a parameter related to the DC component signal.

[0236] For example, the second counting circuit 222 may be configured to perform Fourier transform on the digital signal to obtain a DC component signal.

[0237] Alternatively or additionally, the second counting circuit 222 may be configured to integrate or sum the digital signal within a predetermined time period to obtain a parameter related to the integration result or the summation result and use it as the parameter characterizing the number of photons in the optical signal.

[0238] For more embodiments of the photometer disclosed herein, reference may be made to the applicant's Chinese prior application CN202211510485.X, the contents of which are incorporated herein by reference.

[0239] A second aspect of the present disclosure provides a chemiluminescence analyzer, comprising a sample preparation device, a substrate providing device, a photometric device, and a controller.

[0240] The sample preparation device is used to prepare a sample liquid to be tested from a blood sample to be tested.

[0241] The substrate providing device is used to provide a luminescent substrate.

[0242] The photometric device includes at least a photometer, which is used to detect the luminescent signal generated in the chemiluminescent reaction, wherein the first linear detection range of the photometer is [A1, A2] photon counts / second, and the first linear detection range represents the range of the number of photons output by the photometer. Within the first linear detection range, the intensity of the luminescent signal generated in the chemiluminescent reaction is linearly related to the photon count output by the photometer, wherein A1 is less than or equal to 2000 and A2 is greater than or equal to 10 8 .

[0243] The controller is configured to obtain a test item of the blood sample to be tested. When the test item is a human chorionic gonadotropin test item:

[0244] Controlling the sample preparation device to pre-dilute the blood sample to be tested with a diluent, and mixing the pre-diluted blood sample to be tested with a reaction reagent in a reaction container, so that human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent for human chorionic gonadotropin antibodies with a marker;

[0245] Controlling the substrate providing device to provide the luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested reacts with the luminescent substrate to perform chemiluminescent reaction;

[0246] controlling a luminometer to detect the luminescent signal generated in the chemiluminescent reaction; and

[0247] The detection result of human chorionic gonadotropin in the blood sample to be tested is obtained according to the luminescent signal and the preset calibration model.

[0248] It can be understood that the various embodiments and advantages described for the chemiluminescence analyzer provided in the first aspect of the present disclosure, especially its sample preparation device 110, substrate providing device 120, photometric device 130 and luminescent substrate, are also applicable to the chemiluminescence analyzer provided in the second aspect of the present disclosure.

[0249] In some embodiments, the chemiluminescence analyzer provided in the second aspect of the present disclosure further includes a sample identification device, such as a barcode scanner or a camera, for identifying the test items of the blood sample to be tested and providing the identified test items to the controller.

[0250] In other embodiments, the chemiluminescence analyzer provided in the second aspect of the present disclosure further includes an input device, such as a user interaction interface, for receiving user input indicating the test items of the blood sample to be tested, and the controller is used to receive the user input from the input device to obtain the test items of the blood sample to be tested.

[0251] A third aspect of the present disclosure provides a chemiluminescence analyzer, comprising a sample preparation device, a substrate providing device, a photometric device, and a data processing device.

[0252] The sample preparation device is at least used for: pre-diluting a blood sample for human chorionic gonadotropin detection using a diluent at a preset dilution ratio before performing human chorionic gonadotropin detection, and mixing the pre-diluted blood sample with a reaction reagent in a reaction container so that human chorionic gonadotropin in the pre-diluted blood sample reacts with the reaction reagent to obtain a sample solution to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent for human chorionic gonadotropin antibodies with a marker;

[0253] The substrate providing device is used to provide a luminescent substrate into a reaction container containing a sample liquid to be tested, so that the marker in the sample liquid to be tested and the luminescent substrate undergo a chemiluminescent reaction, wherein the second linear detection range of the luminescent substrate is [B1, B2] photon counts / second, and the second linear detection range represents the range of the intensity of the luminescent signal generated in the chemiluminescent reaction. Within the second linear detection range, the number of markers participating in the chemiluminescent reaction and the intensity of the luminescent signal generated in the chemiluminescent reaction are linearly related, wherein B1 is less than or equal to 3000 and B2 is greater than or equal to 10 8 , and / or, the ratio of B2 to B1 (B2 / B1) is greater than or equal to 30,000;

[0254] A photometric device, the photometric device comprising at least a photometer, the photometer being used to detect the luminescent signal generated in the chemiluminescent reaction;

[0255] The data processing device is used to obtain the detection result of human chorionic gonadotropin in the blood sample to be tested based on the luminescent signal and the preset calibration model.

[0256] It can be understood that the various embodiments and advantages described for the chemiluminescence analyzer provided in the first aspect of the present disclosure, especially its sample preparation device 110, substrate providing device 120, photometric device 130 and luminescent substrate, are also applicable to the chemiluminescence analyzer provided in the third aspect of the present disclosure.

[0257] A fourth aspect of the present disclosure provides a chemiluminescence analyzer, comprising a sample preparation device, a substrate providing device, a photometric device, and a controller.

[0258] The sample preparation device is used to prepare a sample liquid to be tested from a blood sample to be tested.

[0259] The substrate providing device is used to provide a luminescent substrate.

[0260] The photometric device at least includes a photometer, which is used to detect the luminescent signal generated in the chemiluminescent reaction.

[0261] The controller is configured to obtain a test item of the blood sample to be tested. When the test item is a human chorionic gonadotropin test item:

[0262] Controlling the sample preparation device to pre-dilute the blood sample to be tested with a diluent, and mixing the pre-diluted blood sample to be tested with a reaction reagent in a reaction container, so that human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent for human chorionic gonadotropin antibodies with a marker;

[0263] Controlling the substrate providing device to provide the luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested reacts with the luminescent substrate to perform chemiluminescent reaction;

[0264] controlling a luminometer to detect luminescent signals generated in a chemiluminescent reaction;

[0265] Obtaining a detection result of human chorionic gonadotropin in the blood sample to be tested according to the luminescent signal and a preset calibration model;

[0266] Here, the second linear detection range of the luminescent substrate is [B1, B2] photon counts / second. The second linear detection range represents the range of the intensity of the luminescent signal generated in the chemiluminescent reaction. Within the second linear detection range, the number of markers participating in the chemiluminescent reaction is linearly related to the intensity of the luminescent signal generated in the chemiluminescent reaction, wherein B1 is less than or equal to 3000 and B2 is greater than or equal to 10 8 , and / or, the ratio of B2 to B1 B2 / B1 is greater than or equal to 30000.

[0267] It can be understood that the various embodiments and advantages described for the chemiluminescence analyzer provided in the first aspect of the present disclosure, especially its sample preparation device 110, substrate providing device 120, photometric device 130 and luminescent substrate, as well as the various embodiments and advantages described for the chemiluminescence analyzer provided in the second aspect of the present disclosure are also applicable to the chemiluminescence analyzer provided in the fourth aspect of the present disclosure.

[0268] A fifth aspect of the present disclosure provides a chemiluminescence analyzer, comprising a sample preparation device, a substrate providing device, a photometric device, and a data processing device.

[0269] The sample preparation device is used to pre-dilute all blood samples to be tested containing human chorionic gonadotropin as the test substance using a diluent, and mix the pre-diluted blood samples to be tested with a reaction reagent in a reaction container so that the human chorionic gonadotropin in the pre-diluted blood samples to be tested reacts with the reaction reagent to obtain a sample liquid to be tested. The reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent for human chorionic gonadotropin antibodies with a marker.

[0270] The substrate providing device is used to provide the luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested can react with the luminescent substrate to perform chemiluminescent reaction.

[0271] The photometric device includes at least a photometer for detecting a luminescent signal generated in a chemiluminescent reaction. The photometer includes a receiving component and a processing component. The receiving component is configured to receive the light signal generated in the chemiluminescent reaction and convert the light signal into a corresponding electrical signal. The processing component is configured to be electrically connected to the receiving component and receive the electrical signal from the receiving component. The processing component includes a first photon counting module and a second photon counting module. The first photon counting module is configured to detect the number of pulses in the electrical signal using a pulse recognition method to obtain a first photon counting result. The second photon counting module is configured to process the electrical signal to obtain a parameter representing the number of photons in the light signal, and to obtain a second photon counting result based on the parameter representing the number of photons in the light signal and a preset calibration function. The calibration function represents a mapping relationship between the parameter representing the number of photons in the light signal and the photon counting result. The processing component is further configured to obtain a photon count output by the photometer based on the first and second photon counting results, as well as the output photon count.

[0272] The data processing device is used to obtain the detection result of human chorionic gonadotropin in the blood sample to be tested according to the luminescent signal and the preset calibration model.

[0273] It can be understood that the various embodiments and advantages described for the chemiluminescence analyzer provided in the first aspect of the present disclosure, especially its sample preparation device 110, substrate providing device 120, photometric device 130 and luminescent substrate, are also applicable to the chemiluminescence analyzer provided in the fifth aspect of the present disclosure.

[0274] A sixth aspect of the present disclosure provides a chemiluminescence analyzer, comprising a sample preparation device, a substrate providing device, a photometric device, and a controller.

[0275] The sample preparation device is used to prepare a sample liquid to be tested from a blood sample to be tested.

[0276] The substrate providing device is used to provide a luminescent substrate.

[0277] The photometric device includes at least a photometer, which is used to detect the luminescence signal generated in the chemiluminescent reaction. The photometer includes a receiving component and a processing component. The receiving component is configured to receive the light signal generated in the chemiluminescent reaction and convert the light signal into a corresponding electrical signal. The processing component is configured to be electrically connected to the receiving component and receive the electrical signal from the receiving component. The processing component includes a first photon counting module and a second photon counting module. The first photon counting module is configured to detect the number of pulses in the electrical signal using a pulse recognition method to obtain a first photon counting result. The second photon counting module is configured to process the electrical signal to obtain a parameter representing the number of photons in the light signal, and to obtain a second photon counting result based on the parameter representing the number of photons in the light signal and a preset calibration function. The calibration function represents a mapping relationship between the parameter representing the number of photons in the light signal and the photon counting result. The processing component is further configured to obtain a photon count output by the photometer based on the first and second photon counting results, as well as the output photon count.

[0278] The controller is configured to obtain a test item of the blood sample to be tested. When the test item is a human chorionic gonadotropin test item:

[0279] Controlling the sample preparation device to pre-dilute the blood sample to be tested with a diluent, and mixing the pre-diluted blood sample to be tested with a reaction reagent in a reaction container, so that human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample liquid to be tested, wherein the reaction reagent includes a capture reagent coated with human chorionic gonadotropin antibodies and a detection reagent containing human chorionic gonadotropin antibodies with a marker;

[0280] Controlling the substrate providing device to provide the luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested reacts with the luminescent substrate to perform chemiluminescent reaction;

[0281] controlling a luminometer to detect the luminescent signal generated in the chemiluminescent reaction; and

[0282] The detection result of human chorionic gonadotropin in the blood sample to be tested is obtained according to the luminescent signal and the preset calibration model.

[0283] It can be understood that the various embodiments and advantages described for the chemiluminescence analyzer provided in the first aspect of the present disclosure, especially its sample preparation device 110, substrate providing device 120, photometric device 130 and luminescent substrate, as well as the various embodiments and advantages described for the chemiluminescence analyzer provided in the second aspect of the present disclosure are also applicable to the chemiluminescence analyzer provided in the sixth aspect of the present disclosure.

[0284] A seventh aspect of the present disclosure also provides another chemiluminescence analyzer, comprising:

[0285] HCG measuring unit, used to obtain the test result of human chorionic gonadotropin of the blood sample to be tested;

[0286] a mode selection unit, configured to select a first HCG detection mode or a second HCG detection mode; and

[0287] Controller, configured as,

[0288] Mode selection of the receiving mode selection unit,

[0289] When the mode selection unit selects the first HCG detection mode, the HCG measurement unit is controlled to execute a first measurement process, the first measurement process comprising: pre-diluting at least a portion of the blood sample to be tested using a diluent, and performing chemiluminescence measurement on the pre-diluted portion of the blood sample to be tested to obtain and output a first HCG detection result; and

[0290] When the mode selection unit selects the second HCG detection mode, the HCG measurement unit is controlled to execute the second measurement process, which includes: performing chemiluminescence measurement on the undiluted portion of the blood sample to be tested to obtain a second HCG detection result; when the second HCG detection result is abnormal, pre-diluting another portion of the blood sample to be tested with a diluent, and performing chemiluminescence measurement on the pre-diluted portion of the blood sample to be tested to obtain and output a third HCG detection result.

[0291] In some embodiments, the HCG assay unit includes the aforementioned sample preparation device, substrate supply device, and photometry device. For more information on various embodiments and their advantages, please refer to the above description. In other words, the aforementioned features can be applied in corresponding manners to chemiluminescence analyzers with two HCG assay processes.

[0292] In a specific example, the first measurement process includes: the sample preparation device uses a diluent to pre-dilute at least a portion of the blood sample to be tested, and mixes the pre-diluted blood sample portion to be tested with a reaction reagent in a reaction container to obtain a first sample liquid to be tested; the substrate providing device provides a luminescent substrate to the reaction container containing the first sample liquid to be tested, so that the first sample liquid to be tested and the luminescent substrate undergo a chemiluminescent reaction; the photometric device detects the luminescent signal generated in the chemiluminescent reaction to obtain a first HCG detection result. The second measurement process includes: the sample preparation device mixes a portion of the blood sample to be tested with a reaction reagent in a reaction container without dilution to obtain a second sample liquid to be tested; the substrate providing device provides a luminescent substrate to the reaction container containing the second sample liquid to be tested so that the second sample liquid to be tested and the luminescent substrate undergo a chemiluminescent reaction; the photometric device detects the luminescent signal generated in the chemiluminescent reaction to obtain a second HCG test result; when the second HCG test result is abnormal, the sample preparation device uses a diluent to pre-dilute another portion of the blood sample to be tested, and mixes the pre-diluted blood sample portion to be tested with the reaction reagent in the reaction container to obtain a third sample liquid to be tested; the substrate providing device provides the luminescent substrate to the reaction container containing the third sample liquid to be tested so that the third sample liquid to be tested and the luminescent substrate undergo a chemiluminescent reaction; the photometric device detects the luminescent signal generated in the chemiluminescent reaction to obtain a third HCG test result.

[0293] In this embodiment, the chemiluminescence analyzer has two test modes. The first mode is to directly dilute and obtain the actual HCG content based on the test results after dilution. The second mode is to directly test. If the test result exceeds the linear detection range, dilution is performed and retesting is performed. The configuration of two modes allows customers to flexibly choose according to their needs and provide a better user experience.

[0294] An eighth aspect of the present disclosure provides a chemiluminescence analyzer, comprising a sample preparation device, a substrate providing device, a photometric device, a data processing device, and a control device.

[0295] A sample preparation device is used to mix a blood sample to be tested with a reaction reagent in a reaction container so that the substance to be tested in the blood sample to be tested reacts with the reaction reagent to obtain a sample liquid to be tested, wherein the reaction reagent includes a capture reagent and a detection reagent with a marker;

[0296] A reagent carrying device is used to place the test kit, which carries the reaction reagent corresponding to the test item. The test kit corresponding to the human chorionic gonadotropin test item also carries a diluent;

[0297] The substrate providing device is used to provide the luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested and the luminescent substrate can undergo chemiluminescent reaction;

[0298] A photometric device, the photometric device comprising at least a photometer, the photometer being used to detect the luminescent signal generated in the chemiluminescent reaction;

[0299] A data processing device, configured to obtain a detection result of a substance to be tested in a blood sample to be tested based on the luminescence signal and a preset calibration model;

[0300] a control device for controlling the sample preparation device to pre-dilute the blood sample to be tested using a diluent in a test kit corresponding to the human chorionic gonadotropin test item at a preset dilution ratio when the test item of the blood sample to be tested is a human chorionic gonadotropin test item, and to mix the pre-diluted blood sample to be tested with at least a portion of a reaction reagent in the test kit corresponding to the human chorionic gonadotropin test item in a reaction container, so that the human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested;

[0301] Controlling the substrate providing device to provide the luminescent substrate into the reaction container containing the sample liquid to be tested, so that the marker in the sample liquid to be tested reacts with the luminescent substrate to undergo chemiluminescent reaction;

[0302] The photometer is controlled to detect the luminescent signal generated in the chemiluminescent reaction, and the detection result of the human chorionic gonadotropin in the blood sample to be tested is obtained according to the luminescent signal and a preset calibration model.

[0303] It can be understood that the various embodiments and advantages described for the chemiluminescence analyzer provided in the first aspect of the present disclosure, especially its sample preparation device 110, substrate providing device 120, photometric device 130 and luminescent substrate, are also applicable to the chemiluminescence analyzer provided in the eighth aspect of the present disclosure.

[0304] In some embodiments, the kit corresponding to the human chorionic gonadotropin detection project includes a carrier and a first cavity, a second cavity and a third cavity opened on the carrier, the first cavity is used to hold a diluent, the second cavity is used to hold a capture reagent, and the third cavity is used to hold the detection reagent.

[0305] This results in the following benefits:

[0306] a) Automatic operation of HCG dilution on the machine;

[0307] b) The manufacturer provides diluent, which can ensure the accuracy of HCG dilution measurement and avoid poor test results caused by customers preparing diluent on their own;

[0308] c) Save reagent storage space and improve reagent space utilization efficiency;

[0309] It saves testing costs and provides great convenience to customers as they do not need to inventory or purchase diluents separately.

[0310] Those skilled in the art will appreciate that the embodiments of the present disclosure may be provided as methods, systems, or computer program products. Therefore, the embodiments of the present disclosure may take the form of hardware embodiments, software embodiments, or embodiments combining software and hardware. Furthermore, the embodiments of the present disclosure may take the form of a computer program product implemented on one or more computer-usable storage media (including magnetic disk storage and optical storage, etc.) containing computer-usable program code.

[0311] The present disclosure embodiments are described with reference to the flowcharts and / or block diagrams of the methods, devices (systems), and computer program products according to the present disclosure embodiments. It should be understood that each process and / or box in the flowchart and / or block diagram, as well as the combination of the processes and / or boxes in the flowchart and / or block diagram, can be implemented by computer program operations. These computer programs can be provided to operate on a processor of a general-purpose computer, a special-purpose computer, an embedded processor, or other programmable data processing device to produce a machine, so that the operations performed by the processor of the computer or other programmable data processing device produce a device for implementing the functions specified in one or more processes in the flowchart and / or one or more boxes in the block diagram.

[0312] These computer program operations may also be stored in a computer-readable memory that can direct a computer or other programmable data processing device to operate in a specific manner, so that the operations stored in the computer-readable memory produce an article of manufacture including an operating device that implements the functions specified in one or more processes in the flowchart and / or one or more boxes in the block diagram.

[0313] These computer program operations may also be loaded onto a computer or other programmable data processing device so that a series of operational steps are executed on the computer or other programmable device to produce a computer-implemented process, whereby the operations executed on the computer or other programmable device provide steps for implementing the functions specified in one or more processes in the flowchart and / or one or more boxes in the block diagram.

[0314] The features or feature combinations mentioned above in the description, drawings and claims may be used in any combination or alone as long as they are meaningful within the scope of the present disclosure and do not contradict each other.

[0315] The following specific examples illustrate various embodiments and advantages of the chemiluminescent substrate disclosed herein, but the scope of the present disclosure is not limited thereto.

[0316] Example 1: Linear performance using different chemiluminescent substrates

[0317] Chemiluminescent substrate solution formula:

[0318] Chemiluminescent substrate: 200 mg

[0319] 5(6)-Carboxyfluorescein: 200 mg

[0320] Polyvinylbenzyltrimethylammonium chloride: 5g

[0321] AMP-HCl: 50 mM

[0322] Magnesium chloride: 200mg

[0323] Sodium azide: 1g

[0324] Water: Add to a total volume of 1L

[0325] According to the above formula, wherein the chemiluminescent substrate is shown in Table 1 below, a series of chemiluminescent substrate solutions 1-1 to 1-4 were prepared.

[0326] Table 1

[0327] The above-mentioned chemiluminescent substrate solutions 1-1 to 1-4 were tested on a Mindray CL-6000i fully automatic chemiluminescent immunoassay instrument. Alkaline phosphatase (AP) solutions of varying concentration gradients were mixed with each chemiluminescent substrate solution. After incubation for a period of time, light signals were collected. The background signal value (i.e., alkaline phosphatase concentration of 0) and luminescence signal value were obtained at a 2-min illumination time. The alkaline phosphatase concentration in the system and the detection signal values ​​are shown in Table 2 below.

[0328] Table 2

[0329] The light signals (photon counts / second) measured for the chemiluminescent substrate solutions 1-1 to 1-4 were plotted against the concentration of alkaline phosphatase in the system, and linear fitting was performed, as shown in FIG1A to FIG1D .

[0330] According to the data in Table 2 and Figures 16A to 16D, chemiluminescent substrate solutions 1-2, 1-3, and 1-4 have low background signals. Chemiluminescent substrate solutions 1-3 and 1-4 also have high luminescence efficiency and good linearity within the detected luminescence range (R2 ≥ 0.99), among which ADP-STAR performs best.

[0331] Example 2: Linear performance using ADP-STAR and different fluorescent agents

[0332] Chemiluminescent substrate solution formula:

[0333] ADP-STAR: 200mg

[0334] Fluorescent agent: 200mg

[0335] Polyvinylbenzyltrimethylammonium chloride: 5g

[0336] AMP-HCl: 50 mM

[0337] Magnesium chloride: 200mg

[0338] Sodium azide: 1g

[0339] Water: Add to a total volume of 1L

[0340] According to the above formula, wherein the fluorescent agent is shown in Table 3 below, a series of chemiluminescent substrate solutions 2-1 to 2-4 were prepared.

[0341] Table 3

[0342] The above-mentioned chemiluminescent substrate solutions 2-1 to 2-4 were tested on a Mindray CL-6000i fully automatic chemiluminescent immunoassay instrument. Alkaline phosphatase solutions of varying concentration gradients were mixed with each chemiluminescent substrate solution. After incubation for a period of time, light signals were collected. The background signal value (i.e., alkaline phosphatase concentration of 0) and the luminescence signal value were obtained at a 2-min illumination time. The alkaline phosphatase concentration in the system and the detection signal values ​​are shown in Table 4 below.

[0343] Table 4

[0344] The light signals (photon counts / second) measured for the chemiluminescent substrate solutions 2-1 to 2-4 were plotted against the concentration of alkaline phosphatase in the system, and linear fitting was performed, as shown in FIG2A to FIG2D .

[0345] According to the data in Table 4 and Figures 17A to 17D, the chemiluminescent substrate solutions 2-1 to 2-4 all exhibited low background signals and high luminescence efficiency, and had good linearity (R2≥0.99) within the detected luminescence range.

[0346] Example 3: Linear performance using CDP-STAR and different fluorescent agents

[0347] Prepare the chemiluminescent substrate solution according to the following formula:

[0348] CDP-STAR: 200mg

[0349] Fluorescent agent: 200mg

[0350] Polyvinylbenzyltrimethylammonium chloride: 5g

[0351] AMP-HCl: 50 mM

[0352] Magnesium chloride: 200mg

[0353] Sodium azide: 1g

[0354] Water: Add to a total volume of 1L

[0355] According to the above formula, wherein the fluorescent agent is shown in Table 5 below, a series of chemiluminescent substrate solutions 3-1 to 3-4 were prepared.

[0356] Table 5

[0357] The above-mentioned chemiluminescent substrate solutions 3-1 to 3-4 were tested on a Mindray CL-6000i fully automatic chemiluminescent immunoassay instrument. Alkaline phosphatase solutions of varying concentration gradients were mixed with each chemiluminescent substrate solution. After incubation for a period of time, the light signal was collected. The background signal value (i.e., alkaline phosphatase concentration of 0) and the luminescence signal value were obtained at a 2-minute illumination time. The alkaline phosphatase concentration in the system and the detection signal values ​​are shown in Table 6 below.

[0358] Table 6

[0359] The light signals (photon counts / second) measured for the chemiluminescent substrate solutions 3-1 to 3-4 were plotted against the concentration of alkaline phosphatase in the system, and linear fitting was performed, as shown in FIG3A to FIG3D .

[0360] According to the data in Table 6 and Figures 18A to 18D, chemiluminescent substrate solutions 3-1 to 3-4 all exhibit low background signals and high luminescence efficiency, and have good linearity within the detected luminescence range (R2≥0.99).

[0361] Example 4: Linear performance using ADP-STAR and different quaternary ammonium salt cationic surfactants

[0362] Chemiluminescent substrate solution formula:

[0363] ADP-STAR: 200mg

[0364] Fluorescein: 200mg

[0365] Quaternary ammonium surfactant: 5g

[0366] AMP-HCl: 50 mM

[0367] Magnesium chloride: 200mg

[0368] Sodium azide: 1g

[0369] Water: Add to a total volume of 1L

[0370] According to the above formula, wherein the quaternary ammonium salt cationic surfactant is shown in Table 7 below, a series of chemiluminescent substrate solutions 4-1 to 4-4 were prepared.

[0371] Table 7

[0372] The above-mentioned chemiluminescent substrate solutions 4-1 to 4-4 were tested on a Mindray CL-6000i fully automatic chemiluminescent immunoassay instrument. Alkaline phosphatase solutions of varying concentration gradients were mixed with each chemiluminescent substrate solution. After incubation for a period of time, the light signal was collected. The background signal value (i.e., alkaline phosphatase concentration of 0) and the luminescence signal value were obtained at a 2-minute illumination time. The alkaline phosphatase concentration in the system and the detection signal values ​​are shown in Table 8 below.

[0373] Table 8

[0374] The light signals (photon counts / second) measured for the chemiluminescent substrate solutions 4-1 to 4-4 were plotted against the concentration of alkaline phosphatase in the system, and linear fitting was performed, as shown in FIG4A to FIG4D .

[0375] As shown in Table 8 and Figures 19A to 19D , chemiluminescent substrate solutions 4-1 to 4-3 had low background signals and high luminescence efficiencies, exhibiting good linearity within the detected luminescence range (R2 ≥ 0.99). Chemiluminescent substrate solution 4-4 had high background signals and low luminescence efficiency, failing to maintain good linearity within the detected luminescence range (R2 < 0.99), and the ratio of the light signal between the upper and lower detection limits was also small (less than 3000).

[0376] Example 5: Linear performance using CDP-STAR and different quaternary ammonium salt cationic surfactants

[0377] Chemiluminescent substrate solution formula:

[0378] CDP-STAR: 200mg

[0379] Fluorescein: 200mg

[0380] Quaternary ammonium surfactant: 5g

[0381] AMP-HCl: 50 mM

[0382] Magnesium chloride: 200mg

[0383] Sodium azide: 1g

[0384] Water: Add to a total volume of 1L

[0385] According to the above formula, wherein the quaternary ammonium salt cationic surfactant is shown in Table 9 below, a series of chemiluminescent substrate solutions 5-1 to 5-4 were prepared.

[0386] Table 9

[0387] The above-mentioned chemiluminescent substrate solutions 5-1 to 5-4 were tested on a Mindray CL-6000i fully automatic chemiluminescent immunoassay instrument. Alkaline phosphatase solutions of varying concentration gradients were mixed with each chemiluminescent substrate solution. After incubation for a period of time, the light signal was collected. The background signal value (i.e., alkaline phosphatase concentration of 0) and the luminescence signal value were obtained at a 2-minute illumination time. The alkaline phosphatase concentration in the system and the detection signal values ​​are shown in Table 10 below.

[0388] Table 10

[0389] The light signals (photon counts / second) measured for the chemiluminescent substrate solutions 5-1 to 5-4 were plotted against the concentration of alkaline phosphatase in the system, and linear fitting was performed, as shown in FIG5A to FIG5D .

[0390] As shown in the data in Table 10 and Figures 20A to 20D, chemiluminescent substrate solutions 5-1 to 5-3 had low background signals and high luminescence efficiencies, exhibiting good linearity within the detected luminescence range (R2 ≥ 0.99). Chemiluminescent substrate solution 5-4 had high background signals and low luminescence efficiencies, failing to maintain good linearity within the detected luminescence range (R2 < 0.99), and the ratio of the light signal between the upper and lower detection limits was also small (less than 3000).

[0391] Example 6: HCG broad linear detection

[0392] Prepare chemiluminescent substrate solution 6-1 according to the following formula:

[0393] ADP-STAR: 200mg

[0394] Fluorescein: 200mg

[0395] Polyvinylbenzyltrimethylammonium chloride: 5g

[0396] AMP-HCl: 50 mM

[0397] Magnesium chloride: 200mg

[0398] Sodium azide: 1g

[0399] Water: Add to a total volume of 1L

[0400] Prepare chemiluminescent substrate solution 6-2 according to the following formula:

[0401] CDP-STAR: 200mg

[0402] Fluorescein: 200mg

[0403] Polyvinylbenzyltrimethylammonium chloride: 5g

[0404] AMP-HCl: 50 mM

[0405] Magnesium chloride: 200mg

[0406] Sodium azide: 1g

[0407] Water: Add to a total volume of 1L

[0408] Using the aforementioned chemiluminescent substrate solutions 6-1 and 6-2, serum samples containing varying HCG concentrations were tested on a Mindray CL-6000i fully automatic chemiluminescent immunoassay instrument. Sample concentrations ranged from 1 to 200,000 mIU / mL. The HCG detection kit used was the Mindray Total β-Human Chorionic Gonadotropin (Total βHCG) Assay Kit (Chemiluminescent Immunoassay). The photon counts per second of the light signal collected during a 2-minute incubation of each sample with chemiluminescent substrate solutions 6-1 and 6-2 were recorded. The HCG concentration in the sample was calculated based on the light signal and recorded as the back-calculated concentration. The relative deviation of the back-calculated concentration was calculated relative to the nominal HCG concentration in the sample. The results are shown in Tables 11 and 12 below.

[0409] Table 11

[0410] Table 12

[0411] According to Tables 11 and 12, the chemiluminescent substrate solution disclosed herein can be directly detected and analyzed without dilution in the HCG test when the HCG concentration in the sample is in the range of 1 to 200,000 mIU / mL, and the deviation of the test results relative to the nominal concentration is within ±5%.

[0412] The above description is only a preferred embodiment of the present disclosure and does not limit the patent scope of the present disclosure. All equivalent transformations made by using the contents of the present disclosure and the drawings under the inventive concept of the present disclosure, or direct / indirect application in other related technical fields are included in the patent protection scope of the present disclosure.

Claims

1. A chemiluminescence analyzer, characterized in that, Comprising: A sample preparation device, at least for: for a blood sample to be tested for the detection item of human chorionic gonadotropin, before performing the detection of human chorionic gonadotropin on the blood sample to be tested, pre-diluting the blood sample to be tested with a diluent at a preset dilution ratio, and mixing the pre-diluted blood sample to be tested with a reaction reagent in a reaction container, so that human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested, and the reaction reagent includes a capture reagent coated with an antibody against human chorionic gonadotropin and a detection reagent of an antibody against human chorionic gonadotropin with a label; A substrate providing device, for providing a luminescent substrate into the reaction container containing the sample solution to be tested, so that the label in the sample solution to be tested undergoes a chemiluminescence reaction with the luminescent substrate; A photometric device, the photometric device at least includes a photometer, the photometer is used to detect the luminescence signal generated in the chemiluminescence reaction, wherein, the first linear detection range of the photometer is [A1, A2] photon counts per second, the first linear detection range represents the range of photon counts output by the photometer, within this first linear detection range, the intensity of the luminescence signal generated in the chemiluminescence reaction is linearly related to the photon counts output by the photometer, wherein, A1 is less than or equal to 2000 and A2 is greater than or equal to 10 8 ; A data processing device, for obtaining the detection result of human chorionic gonadotropin in the blood sample to be tested according to the luminescence signal and a preset calibration model.

2. The chemiluminescence analyzer according to claim 1, wherein The calibration model is obtained by the chemiluminescence analyzer using a pre-diluted calibrator.

3. The chemiluminescence analyzer according to claim 2, wherein The pre-dilution multiple of the blood sample to be tested is equal to the pre-dilution multiple of the calibrator.

4. The chemiluminescence analyzer according to any one of claims 1 to 3, characterized in that, The diluent, the capture reagent and the detection reagent are placed in a common reagent kit, and the reagent kit includes a carrier and a first cavity, a second cavity and a third cavity opened on the carrier, the first cavity is used to accommodate the diluent, the second cavity is used to accommodate the capture reagent, and the third cavity is used to accommodate the detection reagent.

5. The chemiluminescence analyzer according to any one of claims 1 to 4, characterized in that, The ratio A2 / A1 of A2 to A1 is greater than or equal to 30000.

6. The chemiluminescence analyzer according to any one of claims 1 to 5, characterized in that The second linear detection range of the luminescent substrate is [B1, B2] photon counts per second, and the second linear detection range represents the range of the intensity of the luminescent signal generated in the chemiluminescence reaction. Within this second linear detection range, the number of the labels participating in the chemiluminescence reaction has a linear relationship with the intensity of the luminescent signal generated in the chemiluminescence reaction, where B1 is less than or equal to 3000 and B2 is greater than or equal to 10 8 , and / or, the ratio B2 / B1 of B2 to B1 is greater than or equal to 30000.

7. The chemiluminescence analyzer according to claim 6, wherein Wherein, The lower limit B1 of the second linear detection range is less than or equal to 2000.

8. The chemiluminescence analyzer according to any one of claims 6 to 7, characterized in that, The chemiluminescence analyzer is designed such that: when the concentration of human chorionic gonadotropin in the blood sample to be tested is greater than or equal to 200000 mIU / ml, the intensity of the luminescence signal generated in the chemiluminescence reaction is less than or equal to the smaller value of A2 and B2.

9. The chemiluminescence analyzer according to any one of claims 6 to 8, characterized in that, The chemiluminescence analyzer is designed such that: when the concentration of human chorionic gonadotropin in the blood sample to be tested is greater than or equal to 0.5 mIU / ml, the intensity C1 of the luminescence signal generated in the chemiluminescence reaction is greater than or equal to the larger value of A1 and B1.

10. The chemiluminescence analyzer according to claim 9, characterized in that, The chemiluminescence analyzer is designed such that when the concentration of human chorionic gonadotropin in the blood sample to be tested is greater than 0.5 mIU / ml and less than or equal to 5 mIU / ml, preferably less than or equal to 2 mIU / ml, the intensity of the luminescence signal generated in the chemiluminescence reaction is C2, wherein the ratio C2 / C1 of C2 to C1 is greater than 2.

11. The chemiluminescence analyzer according to any one of claims 6 to 10, characterized in that, The upper limit B2 of the second linear detection range is greater than the upper limit A2 of the first linear detection range.

12. The chemiluminescence analyzer according to any one of claims 1 to 11, characterized in that, The luminescent substrate is designed such that the background of the luminescent substrate is less than 3000; and / or The luminescent substrate is designed such that the signal-to-noise ratio of the luminescent substrate is greater than 40000.

13. The chemiluminescence analyzer according to any one of claims 1 to 12, characterized in that, The luminescent substrate includes a chemiluminescent substrate and a chemiluminescent enhancer. The chemiluminescent enhancer includes a fluorescent agent and a surfactant. The fluorescent agent includes one or more carboxyfluoresceins.

14. The chemiluminescence analyzer according to any one of claims 1 to 13, characterized in that, The luminescent substrate is selected from dioxetane compounds, preferably one or more of AMPPD, CSPD, CDP-STAR, and more preferably CDP-STAR.

15. The chemiluminescence analyzer according to any one of claims 1 to 14, characterized in that The luminescent substrate is selected from a chlorinated derivative of AMPPD, fluorescein or its carboxyl-substituted derivative, and a water-soluble polymeric quaternary ammonium salt cationic surfactant.

16. The chemiluminescence analyzer according to claim 15, wherein The chlorinated derivative of AMPPD is ADP-STAR or CDP-STAR, or The fluorescein or its carboxyl-substituted derivative is fluorescein or 5(6)-carboxyfluorescein, or The water-soluble polymeric quaternary ammonium salt cationic surfactant is polyvinylbenzyltrimethylammonium chloride.

17. The chemiluminescence analyzer according to any one of claims 1 to 16, characterized in that, The photometer includes: a receiving component configured to receive the optical signal generated in the chemiluminescent reaction and convert the optical signal into a corresponding electrical signal; and a processing component configured to be electrically connected to the receiving component and receive the electrical signal from the receiving component. The processing component includes a first photon counting module and a second photon counting module. The first photon counting module is configured to detect the number of pulses of the electrical signal using a pulse recognition method to obtain a first photon counting result. The second photon counting module is configured to process the electrical signal to obtain a parameter characterizing the number of photons in the optical signal, and obtain a second photon counting result according to the parameter characterizing the number of photons in the optical signal and a preset calibration function. Wherein, the calibration function represents the mapping relationship between the parameter characterizing the number of photons in the optical signal and the photon counting result. The processing component is further configured to obtain the photon counting output by the photometer based on the first photon counting result and the second photon counting result, and output the photon counting.

18. The chemiluminescence analyzer according to claim 17, wherein The processing component is further configured to: when the first photon counting result is lower than the first threshold, use the first photon counting result as the photon counting output by the photometer; and when the first photon counting result is not lower than the first threshold, use the second photon counting result as the photon counting output by the photometer.

19. The chemiluminescence analyzer according to claim 17 or 18, characterized in that, The first photon counting module includes at least one first AD conversion circuit and a first counting circuit that are electrically connected to each other. The first AD conversion circuit is configured to collect the electrical signal at a sampling frequency greater than 1 GHz / s and convert the collected electrical signal into a digital signal and output it to the first counting circuit. The first counting circuit is configured to analyze the received digital signal to identify and receive the number of pulses, and then obtain the first photon counting result.

20. The chemiluminescence analyzer according to any one of claims 17 to 19, characterized in that, The second photon counting module is configured to process the electrical signal to obtain a DC component signal, and the parameter characterizing the number of photons in the optical signal includes a parameter related to the DC component signal; preferably, the second photon counting module is configured to integrate the electrical signal within a predetermined time period to obtain an integration result, and the parameter characterizing the number of photons in the optical signal includes a parameter related to the integration result.

21. The chemiluminescence analyzer according to claim 20, characterized in that, The second photon counting module includes a second AD conversion circuit and a second counting circuit that are electrically connected to each other. The second AD conversion circuit is configured to convert the electrical signal into a digital signal. The second counting circuit is configured to process the digital signal to obtain a parameter characterizing the number of photons in the optical signal, and obtain the second photon counting result according to the parameter characterizing the number of photons in the optical signal and a preset calibration function; preferably, the second counting circuit is configured to integrate or sum the digital signal within a predetermined time period to obtain a parameter related to the integration result or the summation result and use it as the parameter characterizing the number of photons in the optical signal.

22. The chemiluminescence analyzer according to any one of claims 1 to 21, characterized in that, The preset dilution ratio includes 2 to 20 times.

23. A chemiluminescence analyzer, characterized in that, Comprising: A sample preparation device for preparing a sample solution to be tested from a blood sample to be tested; A substrate providing device for providing a luminescent substrate; A photometric device, the photometric device at least includes a photometer, the photometer is used to detect the luminescence signal generated in the chemiluminescence reaction, wherein, the first linear detection range of the photometer is [A1, A2] photon counts per second, the first linear detection range represents the range of the output photon counts of the photometer, within this first linear detection range, the intensity of the luminescence signal generated in the chemiluminescence reaction is linearly related to the photon counts output by the photometer, wherein, A1 is less than or equal to 2000 and A2 is greater than or equal to 10 8 ; A controller configured to Obtain the test items of the blood sample to be tested, When the test item is a human chorionic gonadotropin test item: Control the sample preparation device to pre-dilute the blood sample to be tested with a diluent, and mix the pre-diluted blood sample to be tested with a reaction reagent in a reaction container, so that the human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested. The reaction reagent includes a capture reagent coated with a human chorionic gonadotropin antibody and a detection reagent with a labeled human chorionic gonadotropin antibody. Control the substrate providing device to provide the luminescent substrate into the reaction container containing the sample solution to be tested, so that the label in the sample solution to be tested undergoes a chemiluminescence reaction with the luminescent substrate. Control the photometer to detect the luminescence signal generated in the chemiluminescence reaction. Obtain the test result of the human chorionic gonadotropin in the blood sample to be tested according to the luminescence signal and a preset calibration model.

24. A chemiluminescence analyzer, characterized in that, Comprising: A sample preparation device, at least for: for a blood sample to be tested with a human chorionic gonadotropin test item, before performing the human chorionic gonadotropin test, pre-dilute the blood sample to be tested with a diluent at a preset dilution ratio, and mix the pre-diluted blood sample to be tested with a reaction reagent in a reaction container, so that the human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested. The reaction reagent includes a capture reagent coated with a human chorionic gonadotropin antibody and a detection reagent with a labeled human chorionic gonadotropin antibody; A substrate providing device is used to provide a luminescent substrate into the reaction vessel containing the sample solution to be tested, so that the marker in the sample solution to be tested undergoes a chemiluminescence reaction with the luminescent substrate. Wherein, the second linear detection range of the luminescent substrate is [B1, B2] photon counts per second, and the second linear detection range represents the range of the intensity of the luminescence signal generated in the chemiluminescence reaction. Within the second linear detection range, the number of the markers participating in the chemiluminescence reaction is linearly related to the intensity of the luminescence signal generated in the chemiluminescence reaction, where B1 is less than or equal to 3000 and B2 is greater than or equal to 10 8 , and / or, the ratio B2 / B1 of B2 to B1 is greater than or equal to 30000; A photometric device, the photometric device at least includes a photometer, and the photometer is used to detect the luminescence signal generated in the chemiluminescence reaction; A data processing device, which is used to obtain the detection result of human chorionic gonadotropin in the blood sample to be tested according to the luminescence signal and a preset calibration model.

25. A chemiluminescence analyzer, characterized in that, Comprising: A sample preparation device, which is used to prepare a sample solution to be tested from a blood sample to be tested; A substrate providing device, which is used to provide a luminescent substrate; A photometric device, the photometric device at least includes a photometer, and the photometer is used to detect the luminescence signal generated in the chemiluminescence reaction; A controller, configured to Obtain the test items of the blood sample to be tested, When the test item is a human chorionic gonadotropin test item: Control the sample preparation device to pre-dilute the blood sample to be tested with a diluent, and mix the pre-diluted blood sample to be tested with a reaction reagent in a reaction container, so that the human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested. The reaction reagent includes a capture reagent coated with a human chorionic gonadotropin antibody and a detection reagent with a labeled human chorionic gonadotropin antibody; Control the substrate providing device to provide the luminescent substrate into the reaction container containing the sample solution to be tested, so that the label in the sample solution to be tested undergoes a chemiluminescence reaction with the luminescent substrate; Control the photometer to detect the luminescence signal generated in the chemiluminescence reaction, Obtain the detection result of human chorionic gonadotropin in the blood sample to be tested according to the luminescence signal and a preset calibration model, Among them, the second linear detection range of the luminescent substrate is [B1, B2] photon counts per second, and the second linear detection range represents the range of the intensity of the luminescent signal generated in the chemiluminescence reaction. Within the second linear detection range, the number of the markers participating in the chemiluminescence reaction has a linear relationship with the intensity of the luminescent signal generated in the chemiluminescence reaction, where B1 is less than or equal to 3000 and B2 is greater than or equal to 10 8 , and / or, the ratio B2 / B1 of B2 to B1 is greater than or equal to 30000.

26. A chemiluminescence analyzer, characterized in that, Comprising: A sample preparation device, which is used to pre-dilute all blood samples to be tested containing human chorionic gonadotropin as the substance to be tested with a diluent, and mix the pre-diluted blood samples to be tested with a reaction reagent in a reaction container, so that the human chorionic gonadotropin in the pre-diluted blood samples to be tested reacts with the reaction reagent to obtain a sample solution to be tested. The reaction reagent includes a capture reagent coated with a human chorionic gonadotropin antibody and a detection reagent with a labeled human chorionic gonadotropin antibody; A substrate providing device, which is used to provide the luminescent substrate into the reaction container containing the sample solution to be tested, so that the label in the sample solution to be tested undergoes a chemiluminescence reaction with the luminescent substrate; A photometric device, the photometric device at least includes a photometer, the photometer is used to detect the luminescence signal generated in the chemiluminescence reaction, the photometer includes a receiving component and a processing component, the receiving component is configured to receive the optical signal generated in the chemiluminescence reaction and convert the optical signal into a corresponding electrical signal, the processing component is configured to be electrically connected to the receiving component and receive the electrical signal from the receiving component, the processing component includes a first photon counting module and a second photon counting module, the first photon counting module is configured to detect the number of pulses of the electrical signal by using the pulse recognition method to obtain a first photon counting result, the second photon counting module is configured to process the electrical signal to obtain a parameter characterizing the number of photons in the optical signal, and obtain a second photon counting result according to the parameter characterizing the number of photons in the optical signal and a preset calibration function, wherein, the calibration function represents the mapping relationship between the parameter characterizing the number of photons in the optical signal and the photon counting result, the processing component is further configured to obtain the photon counting output by the photometer based on the first photon counting result and the second photon counting result, and output the photon counting; A data processing device, configured to obtain the detection result of human chorionic gonadotropin in the blood sample to be tested according to the luminescence signal and a preset calibration model.

27. A chemiluminescence analyzer, characterized in that, Comprising: A sample preparation device, configured to prepare a sample solution to be tested from a blood sample to be tested; A substrate providing device, configured to provide a luminescent substrate; A photometric device, the photometric device at least includes a photometer, the photometer is used to detect the luminescence signal generated in the chemiluminescence reaction, the photometer includes a receiving component and a processing component, the receiving component is configured to receive the optical signal generated in the chemiluminescence reaction and convert the optical signal into a corresponding electrical signal, the processing component is configured to be electrically connected to the receiving component and receive the electrical signal from the receiving component, the processing component includes a first photon counting module and a second photon counting module, the first photon counting module is configured to detect the number of pulses of the electrical signal by using the pulse recognition method to obtain a first photon counting result, the second photon counting module is configured to process the electrical signal to obtain a parameter characterizing the number of photons in the optical signal, and obtain a second photon counting result according to the parameter characterizing the number of photons in the optical signal and a preset calibration function, wherein, the calibration function represents the mapping relationship between the parameter characterizing the number of photons in the optical signal and the photon counting result, the processing component is further configured to obtain the photon counting output by the photometer based on the first photon counting result and the second photon counting result, and output the photon counting; A controller, configured to Obtain the test item of the blood sample to be tested, When the test item is a human chorionic gonadotropin test item: Control the sample preparation device to pre-dilute the blood sample to be tested with a diluent, and mix the pre-diluted blood sample to be tested with a reaction reagent in a reaction container, so that human chorionic gonadotropin in the pre-diluted blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested. The reaction reagent includes a capture reagent coated with an antibody against human chorionic gonadotropin and a detection reagent of an antibody against human chorionic gonadotropin with a label. Control the substrate providing device to provide a luminescent substrate into the reaction container containing the sample solution to be tested, so that the label in the sample solution to be tested undergoes a chemiluminescence reaction with the luminescent substrate. Control the photometer to detect the luminescence signal generated in the chemiluminescence reaction. Obtain the detection result of human chorionic gonadotropin in the blood sample to be tested according to the luminescence signal and a preset calibration model.

28. A chemiluminescence analyzer, comprising: an HCG determination unit for obtaining the detection result of human chorionic gonadotropin in a blood sample to be tested; a mode selection unit for selecting a first HCG detection mode or a second HCG detection mode; and a controller configured to receive the mode selection of the mode selection unit, when the mode selection unit selects the first HCG detection mode, control the HCG determination unit to execute a first determination process. The first determination process includes: pre-diluting at least a part of the blood sample to be tested with a diluent, and performing chemiluminescence determination on the pre-diluted part of the blood sample to be tested to obtain and output a first HCG detection result; and when the mode selection unit selects the second HCG detection mode, control the HCG determination unit to execute a second determination process. The second determination process includes: performing chemiluminescence determination on the undiluted part of the blood sample to be tested to obtain a second HCG detection result. When the second HCG detection result is abnormal, pre-dilute another part of the blood sample to be tested with a diluent, and perform chemiluminescence determination on the pre-diluted part of the blood sample to be tested to obtain and output a third HCG detection result.

29. A chemiluminescence analyzer, characterized in that, Comprising: a sample preparation device for mixing a blood sample to be tested with a reaction reagent in a reaction container, so that a substance to be tested in the blood sample to be tested reacts with the reaction reagent to obtain a sample solution to be tested. The reaction reagent includes a capture reagent and a detection reagent with a label; a reagent carrying device for placing a reagent kit. The reagent kit carries reaction reagents corresponding to detection items, and the reagent kit corresponding to the human chorionic gonadotropin detection item also carries a diluent; a substrate providing device for providing a luminescent substrate into the reaction container containing the sample solution to be tested, so that the label in the sample solution to be tested undergoes a chemiluminescence reaction with the luminescent substrate; a light measuring device, the light measuring device at least includes a photometer, and the photometer is used for detecting the luminescence signal generated in the chemiluminescence reaction. A data processing device, configured to obtain a detection result of the substance to be measured in the blood sample to be measured according to the luminescence signal and a preset calibration model; A control device, configured to, when the detection item of the blood sample to be measured is a human chorionic gonadotropin detection item, control the sample preparation device to pre-dilute the blood sample to be measured with a diluent in a kit corresponding to the human chorionic gonadotropin detection item at a preset dilution ratio, and mix at least part of the reaction reagents in the kit corresponding to the human chorionic gonadotropin detection item with the pre-diluted blood sample to be measured in the reaction container, so that the human chorionic gonadotropin in the pre-diluted blood sample to be measured reacts with the reaction reagent to obtain a sample solution to be measured; Control the substrate providing device to provide a luminescent substrate into the reaction container containing the sample solution to be measured, so that the label in the sample solution to be measured undergoes a chemiluminescence reaction with the luminescent substrate; Control the photometer to detect the luminescence signal generated in the chemiluminescence reaction, and obtain the detection result of the human chorionic gonadotropin in the blood sample to be measured according to the luminescence signal and a preset calibration model.

30. The chemiluminescence analyzer according to claim 1 or 29, characterized in that, The kit corresponding to the human chorionic gonadotropin detection item includes a carrier and a first cavity, a second cavity and a third cavity opened on the carrier, the first cavity is used to accommodate the diluent, the second cavity is used to accommodate the capture reagent, and the third cavity is used to accommodate the detection reagent.

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