Quantification method and apparatus for detecting the prozone phenomenon in a sample to be detected - Patents.com

Abstract

To provide a quantification method and device for detecting a prozone phenomenon of a sample awaiting detection.SOLUTION: A method includes the steps of: constructing a first calibration curve by using the actual antigen concentration of a reference sample and the curvature or length of a reaction curve; determining whether or not a sample awaiting detection has a prozone phenomenon on the basis of the curvature or length of the calculated reaction curve of the sample awaiting detection; and when it is determined that the sample awaiting detection has the prozone phenomenon, if the antigen concentration of the sample awaiting detection determined from the first calibration curve that has been constructed is one value, determining the determined antigen concentration of the sample awaiting detection as the reference value of the antigen concentration of the sample awaiting detection, if the determined antigen concentration of the sample awaiting detection is two values, verifying the determined antigen concentration of the sample awaiting detection and determining the antigen concentration of the sample awaiting detection succeeded in verification as the reference value of the antigen concentration of the sample awaiting detection. The present invention can provide a qualitative prompt and a reference value to the sample awaiting detection with the prozone phenomenon.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to the field of detection technology, and in particular to a quantification method and device for detecting the prozone phenomenon in a sample to be detected. [Background technology]

[0002] Immunoturbidimetry is a routine detection method used in fully automatic biochemical analyzers, fully automatic blood coagulation measuring devices and special protein measuring devices, but this method may cause a phenomenon in which the antigen or antibody to be detected becomes excessive during use, and the resulting situation is called the prozone phenomenon, and such a specimen is called a sample with the prozone phenomenon. When detecting a sample with the prozone phenomenon, the device prompts the operator to dilute the sample with the prozone phenomenon before detection and uses several judgment methods to obtain the correct concentration value.

[0003] The method of determining prozone samples used in the current device is based on the change in the slope (i.e., first derivative) of the reaction curve, and selects two time intervals, an unstable interval and a linear interval, on the time axis representing the abscissa, and distinguishes samples with prozone phenomenon based on the fact that the ratio of the slope of the linear interval of the prozone sample to the slope of the unstable interval is greater than that of the normal sample, and the system can further give a prompt for the operator to dilute the sample with prozone phenomenon and provide correct sample concentration information. The method used in the fully automatic biochemical analyzer differs from the above methods in details, but both are methods based on comparing the slope changes of the front and rear segments of the reaction curve. However, the currently used prozone alarm methods are all qualitative, i.e., they are only for determining whether the sample has prozone phenomenon or not, but no method is provided that can quantitatively alarm.

[0004] The above description of the background art is only intended to facilitate a deeper understanding of the technical solutions of the present invention (in terms of the technical means used, the technical problems solved, and the technical effects produced, etc.), and should not be regarded as an admission or in any way suggestion that this information constitutes prior art already known to those skilled in the art. Summary of the Invention

[0005] In response to the shortcomings of the prior art, the present invention provides a quantification method and apparatus for detecting the prozone phenomenon in a sample to be detected, which can provide a qualitative prompt for the sample to be detected that has the prozone phenomenon, and can further provide a reference value for the antigen concentration depending on the length or curvature of the reaction curve.

[0006] According to an embodiment of the present invention, a quantification method for detecting the prozone phenomenon in a sample to be detected is provided, the method comprising the steps of constructing a first calibration curve showing the relationship between the curvature of the reaction curve and the actual antigen concentration, or a second calibration curve showing the relationship between the length of the reaction curve and the actual antigen concentration, using the actual antigen concentration and the curvature or length of the reaction curve of the calculated change in absorbance over time of each of a plurality of reference samples, including a sample with the prozone phenomenon and a sample without the prozone phenomenon, and determining whether the sample to be detected has the prozone phenomenon based on the calculated curvature or length of the reaction curve of the sample to be detected. and if it is determined that the sample to be detected has a prozone phenomenon, determining an antigen concentration of the sample to be detected corresponding to a curvature or length of the reaction curve of the sample to be detected according to the constructed first calibration curve, and if the determined antigen concentration of the sample to be detected is one value, determining the determined antigen concentration of the sample to be detected as a reference value of the antigen concentration of the sample to be detected, and if the determined antigen concentration of the sample to be detected is two values, verifying the determined antigen concentration of the sample to be detected and determining the successfully verified antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected.

[0007] Preferably, a second calibration curve showing the relationship between absorbance and actual antigen concentration is constructed using the absorbance and actual antigen concentration of each of a plurality of reference samples, and if it is determined that there is no prozone phenomenon in the sample to be detected, the antigen concentration of the sample to be detected corresponding to the absorbance of the sample to be detected is determined according to the constructed second calibration curve, and the determined antigen concentration of the sample to be detected is determined as the antigen concentration of the sample to be detected.

[0008] Preferably, when the determined antigen concentration of the sample to be detected has two values, the step of verifying the determined antigen concentration of the sample to be detected and determining the antigen concentration of the sample to be detected that is successfully verified as the reference value of the antigen concentration of the sample to be detected includes the step of verifying the lower value of the determined antigen concentrations of the sample to be detected, and if the verification is successful, determining the lower antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected, and if the verification is unsuccessful, determining the higher antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected.

[0009] Preferably, the step of verifying the lower value of the determined antigen concentrations of the sample to be detected includes the steps of diluting the sample to be detected so that the sample to be detected is a sample free of the prozone phenomenon, and estimating the antigen concentration of the diluted sample to be detected based on the lower value of the antigen concentrations of the sample to be detected; determining the antigen concentration of the diluted sample to be detected using a second calibration curve based on the absorbance of the diluted sample to be detected; and comparing the estimated antigen concentration of the diluted sample to be detected with the determined antigen concentration of the diluted sample to be detected, indicating that the verification of the lower antigen concentration of the sample to be detected has been successful if the estimated antigen concentration of the diluted sample to be detected matches the determined antigen concentration of the diluted sample to be detected; and indicating that the verification of the lower antigen concentration of the sample to be detected has failed if the estimated antigen concentration of the diluted sample to be detected does not match the determined antigen concentration of the diluted sample to be detected.

[0010] Preferably, the step of calculating the curvature or length of the reaction curve of the absorbance over time includes the steps of: collecting data of the transmitted light intensity over time using an optical detection system in the reaction process, converting the data of the transmitted light intensity over time into a reaction curve of the absorbance over time, and determining the absorbance corresponding to a preset time point according to the reaction curve of the absorbance over time; and selecting a preset start time point and a preset end time point on the reaction curve of the absorbance over time, and calculating the curvature or length of the reaction curve of the absorbance over time between the preset start time point and the preset end time point.

[0011] According to another embodiment of the present invention, a quantification device for detecting the prozone phenomenon in a sample to be detected is provided, the device including a first curve construction module, a determination module, a first quantification module, and a second quantification module. The first curve construction module is configured to construct a first calibration curve showing the relationship between the curvature of the reaction curve and the actual antigen concentration, or a first calibration curve showing the relationship between the length of the reaction curve and the actual antigen concentration, using the actual antigen concentration and the curvature or length of the reaction curve of the calculated change in absorbance over time of each of a plurality of reference samples including a sample with the prozone phenomenon and a sample without the prozone phenomenon. The determination module is configured to determine whether the sample to be detected has the prozone phenomenon or not based on the calculated curvature or length of the reaction curve of the sample to be detected. The first quantification module is configured to, when it determines that the sample to be detected has a prozone phenomenon, determine an antigen concentration of the sample to be detected corresponding to a curvature or length of the reaction curve of the sample to be detected according to the constructed first calibration curve, when the determined antigen concentration of the sample to be detected is one value, determine the determined antigen concentration of the sample to be detected as a reference value of the antigen concentration of the sample to be detected, and when the determined antigen concentration of the sample to be detected is two values, verify the determined antigen concentration of the sample to be detected, and determine the successfully verified antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected.

[0012] Preferably, the curve construction module is further configured to construct a second calibration curve showing the relationship between the absorbance and the actual antigen concentration using the absorbance and the actual antigen concentration of each of a plurality of reference samples, and the apparatus further includes a second curve construction module, and the second quantification module is configured to, when it determines that the sample to be detected does not have a prozone phenomenon, determine the antigen concentration of the sample to be detected corresponding to the absorbance of the sample to be detected according to the constructed second calibration curve, and determine the determined antigen concentration of the sample to be detected as the antigen concentration of the sample to be detected.

[0013] Preferably, the first quantification module is configured to verify the lower of the determined antigen concentrations of the sample to be detected, and if the verification is successful, to determine the lower antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected, and if the verification is unsuccessful, to determine the higher antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected.

[0014] Preferably, the first quantification module is configured to, when verifying the lower value of the determined antigen concentrations of the samples to be detected, dilute the sample to be detected so that the sample to be detected is a sample free of the prozone phenomenon, estimate the antigen concentration of the diluted sample to be detected based on the lower value of the antigen concentrations of the samples to be detected, determine the antigen concentration of the diluted sample to be detected using a second calibration curve based on the absorbance of the diluted sample to be detected, compare the estimated antigen concentration of the diluted sample to be detected with the determined antigen concentration of the diluted sample to be detected, and indicate successful verification of the lower antigen concentration of the sample to be detected if the estimated antigen concentration of the diluted sample to be detected matches the determined antigen concentration of the diluted sample to be detected, and indicate unsuccessful verification of the lower antigen concentration of the sample to be detected if the estimated antigen concentration of the diluted sample to be detected does not match the determined antigen concentration of the diluted sample to be detected.

[0015] Preferably, the apparatus includes a calculation module for calculating a curvature or a length of a reaction curve of a time-dependent change in absorbance, the calculation module being configured to collect data of a time-dependent change in transmitted light intensity using an optical detection system in a reaction process, convert the data of a time-dependent change in transmitted light intensity into a reaction curve of a time-dependent change in absorbance, determine an absorbance corresponding to a preset time point according to the reaction curve of a time-dependent change in absorbance, select a preset start time point and a preset end time point on the reaction curve of a time-dependent change in absorbance, and calculate a curvature or a length of the reaction curve of a time-dependent change in absorbance between the preset start time point and the preset end time point.

[0016] The present invention adopts the above technical solution and has the following beneficial effects:

[0017] The present invention can provide a qualitative prompt for samples waiting to be detected that have the prozone phenomenon, and can also provide a reference value for antigen concentration depending on the length or curvature of the reaction curve.

[0018] Exemplary embodiments of the present invention are explained in more detail below in conjunction with the drawings. For the sake of clarity, the same elements in different drawings are designated with the same symbols. As should be understood, the drawings are for schematic purposes only and are not necessarily drawn to scale. These drawings are as follows: [Brief description of the drawings]

[0019] [Figure 1] FIG. 1 is a schematic diagram illustrating a quantification method for detecting the prozone phenomenon in a sample to be detected according to an embodiment of the present invention. [Diagram 2] 1 is a calibration curve showing an example of the relationship between the curvature of the reaction curve and the measured antigen concentration. [Diagram 3] 1 is an exemplary calibration curve showing the relationship between reaction curve length and measured antigen concentration. [Figure 4] FIG. 1 is a block diagram showing the configuration of a quantification device for detecting the prozone phenomenon in a sample to be detected according to an embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] The following describes in detail an embodiment of the present invention, which is implemented on the basis of the technical solution of the present invention, and provides detailed embodiments and specific operation processes, but the protection scope of the present invention is not limited to the following embodiments.

[0021] 1 is a schematic diagram showing a quantification method for detecting the prozone phenomenon in a sample to be detected according to an embodiment of the present invention. As shown in FIG. 1, the quantification method for detecting the prozone phenomenon in a sample to be detected according to an embodiment of the present invention can include the following steps:

[0022] In S100, a first calibration curve showing the relationship between the curvature of the reaction curve and the actual antigen concentration, or a first calibration curve showing the relationship between the length of the reaction curve and the actual antigen concentration, is constructed using the actual antigen concentration of each of the multiple reference samples and the calculated curvature or length of the reaction curve of the change in absorbance over time, the multiple reference samples including a sample with the prozone phenomenon and a sample without the prozone phenomenon. A second calibration curve showing the relationship between the absorbance and the actual antigen concentration is constructed using the absorbance and the actual antigen concentration of each of the multiple reference samples.

[0023] In S200, it is determined whether the sample to be detected has a prozone phenomenon based on the calculated curvature or length of the response curve of the sample to be detected.

[0024] If it is determined that the sample to be detected does not have the prozone phenomenon ("No" in S200), the antigen concentration of the sample to be detected corresponding to the absorbance of the sample to be detected is determined according to the constructed second calibration curve, and the determined antigen concentration of the sample to be detected is determined as the antigen concentration of the sample to be detected (S300).

[0025] Conversely, if it is determined that the sample to be detected contains the prozone phenomenon ("Yes" in S200), the antigen concentration of the sample to be detected corresponding to the curvature or length of the reaction curve of the sample to be detected is determined according to the constructed first calibration curve (S400).

[0026] Next, it is determined whether the antigen concentration of the determined sample to be detected is one value or two values ​​(S500). If the antigen concentration of the determined sample to be detected is one value ("Yes" in S500), the antigen concentration of the determined sample to be detected is determined as the reference value of the antigen concentration of the sample to be detected (S600). If the antigen concentration of the determined sample to be detected is two values ​​("No" in S500), the antigen concentration of the determined sample to be detected is verified, specifically, the lower value of the determined antigen concentrations of the samples to be detected is verified, and if the verification is successful, the lower antigen concentration of the sample to be detected is determined as the reference value of the antigen concentration of the sample to be detected, and if the verification is unsuccessful, the higher antigen concentration of the sample to be detected is determined as the reference value of the antigen concentration of the sample to be detected (S700). The reference value of the antigen concentration of the sample to be detected is used to provide a reference for the actual antigen concentration of the sample to be detected.

[0027] In S100, carcinoembryonic antigen (CEA) is used as an example of sample reagent, the normal range of CEA is 0-5ng / ml, and the linear range is 0-100ng / ml. After diluting the pure CEA in different proportions, 15 standards with different concentrations are obtained. Then, these 15 standards with different concentrations are used as multiple reference samples according to the embodiment of the present invention.

[0028] The actual antigen concentrations of these 15 standards after diluting CEA at different ratios are known, and the time-dependent change data of the transmitted light intensity of the 15 standards is obtained by measuring using an optical detection system, and 15 reaction curves of the time-dependent change of absorbance corresponding to the 15 standards are obtained, and the curvature and length of each reaction curve can be calculated.

[0029] In one exemplary embodiment, the optical detection system is used to illuminate the sample to be detected, and is composed of a light source, a lens, a filter, and an optical fiber, and is located on one side of the analytical instrument, the light emitted from the light source is illuminated onto the analytical instrument, the sample to be detected undergoing a reaction is placed in the analytical instrument, the light passing through the analytical instrument is illuminated onto a receiver, and a signal collecting circuit in the receiver converts the amount of received light into transmitted light intensity, thereby collecting data on the transmitted light intensity over time, thereby forming original signal response data.Then, the original signal response data of the sample to be detected is converted into absorbance over time data, thereby determining a response curve of the change over time of the sample to be detected.

[0030] Because the physical vibration during mixing of the sample and the reagent causes the absorbance data in the initial time interval (e.g., from 0 s to 10 s) to change significantly, the data after a certain period (e.g., 11 s and thereafter) is truly caused by the biochemical reaction, and therefore a preset start time point and a preset end time point are selected on the reaction curve of the absorbance change over time, and the selected preset start time point may be 11 s, and the selected preset end time point may be 27 s. After selecting the preset start time point and the preset end time point on the reaction curve of the absorbance change over time, the curvature or length of the reaction curve of the absorbance change over time between the preset start time point and the preset end time point is calculated.

[0031] When calculating the curvature, a three-point curvature calculation method can be used. Specifically, data between a preset start time point and a preset end time point are fitted according to a preset function model to obtain an arc curve relating to absorbance and time. A first preset time point (e.g., 12 s), a second preset time point (e.g., 19 s) and a third preset time point (e.g., 25 s) are selected on the fitted arc curve. The radius of the fitted arc curve is calculated based on the coordinates of points corresponding to the first preset time point, the second preset time point and the third preset time point on the fitted arc curve, respectively. The reciprocal of the radius is used as the curvature of the fitted arc curve.

[0032] Similarly, below, the curvature or length of the response curve of a sample to be detected is calculated based on the above method.

[0033] Therefore, the curvature, length, measured antigen concentration and actual antigen concentration of the reaction curve for each of the 15 standards can be obtained as shown in Table 1 below.

[0034] [Table 1]

[0035] As shown in Table 1, since the linear range of CEA is 0-100 ng / ml, among the 15 standards, the standards with actual antigen concentrations lower than 100 ng / ml are samples without prozone phenomenon, and the standards with actual antigen concentrations higher than 100 ng / ml are samples with prozone phenomenon, that is, standards No. 1-No. 7 are reference samples without prozone phenomenon, and standards No. 8-No. 15 are reference samples with prozone phenomenon.

[0036] As can be seen from Table 1, the known coordinate points for the actual antigen concentration and curvature of the reaction curve include (3.15,0.0115), (7.76,0.0525), (16.38,0.1018), (25.95,0.1460), (35.54,0.1659), (45.66,0.1999), (83.3,0.2645), (180,0.9000), (360,2.3000), (720,3.2300), (900,3.4280), (1800,3.3260), (3600,3.1096), (7200,2.8813), and (9000,2.7950). According to an embodiment of the present invention, the known coordinate points of these limited points are used to construct a first calibration curve showing the relationship between the curvature of the reaction curve and the actual antigen concentration, and such a process is called a scaling process, and in the scaling process, known scaling function models such as the Logit-Log 3P model, the logit-log 4P model, the logit-log 5P model, etc., can be used. Figure 2 is a first calibration curve exemplarily showing the relationship between the curvature of the reaction curve and the measured antigen concentration, where the X-axis represents the antigen concentration in ng / ml, and the Y-axis represents the curvature in m^-1.

[0037] Similarly, as can be seen from Table 1, the known coordinate points for the actual antigen concentration and reaction curve length include (3.15,0.0080), (7.76,0.0170), (16.38,0.0334), (25.95,0.0505), (35.54,0.0675), (45.66,0.0883), (83.3,0.1526), ​​(180,0.2544), (360,0.3162), (720,0.3156), (900,0.3095), (1800,0.2755), (3600,0.2512), (7200,0.2279), and (9000,0.2182). Using the known coordinates of these limited points, a first calibration curve showing the relationship between the length of the reaction curve and the measured antigen concentration is constructed. Figure 3 shows an example of the first calibration curve showing the relationship between the length of the reaction curve and the actual antigen concentration, where the X-axis represents the antigen concentration in ng / ml and the Y-axis represents the length in m.

[0038] In S200, for example, the monotonic interval (and monotonic stepwise increase) of the first calibration curve showing the relationship between the curvature of the reaction curve and the measured antigen concentration is [0, 900] as shown in Fig. 2. As shown in Fig. 3, the monotonic interval of the first calibration curve showing the relationship between the length of the reaction curve and the measured antigen concentration is [0, 360].

[0039] As can be seen from FIG. 2 and FIG. 3, the monotonic interval [0,900] of the first calibration curve shown in FIG. 2 is wider than the monotonic curve [0,360] of the first calibration curve shown in FIG. 3. Therefore, for the reagent CEA, the curvature of the reaction curve is more sensitive to the prozone phenomenon. In the following, the curvature of the reaction curve is selected as an indicator of the prozone quantification alarm, and the quantification method for detecting the prozone phenomenon of the sample to be detected according to the embodiment of the present invention is described in detail. However, for other reagents, the length of the reaction curve may be more sensitive to the prozone phenomenon, and in this case, the length of the reaction curve can be selected as an indicator of the prozone quantification alarm.

[0040] For the reference samples No. 1 to No. 7 without prozone phenomenon, the absorbance corresponding to a preset time point is determined according to the reaction curve of the change in absorbance over time, and the absorbance and actual antigen concentration of each of the reference samples No. 1 to No. 7 are used to construct a second calibration curve showing the relationship between absorbance and actual antigen concentration. When the antigen binds to a specific antibody in the system, the turbidity of the system increases due to the increase in particles, resulting in an increase in absorbance, so the concentration of the antigen, which is the analyte component in the system, can be estimated by the Lambert-Beer law, and since the Lambert-Beer law shows a direct proportional relationship between absorbance and actual antigen concentration, the second calibration curve is a straight line passing through the origin.

[0041] In S200, it is determined whether the sample to be detected has a prozone phenomenon based on the calculated curvature of the response curve of the sample to be detected. Specifically, a reference curvature can be determined:

[0042] [ka] where K0 is the reference curvature,

[0043] [ka] is the average value of the curvature between a preset start time point and a preset end time point of the reaction curve of the time-dependent change in absorbance of multiple samples without prozone phenomenon, and C is a coefficient, which may be determined by multiple experiments, and its value ranges from 0.8 to 1.2.

[0044] As an example, the reference curvature can be calculated using reference samples No. 1 to No. 7 that are free of the prozone phenomenon. The calculated curvature of the reaction curve of the sample awaiting detection is compared with the reference curvature, and if the calculated curvature of the reaction curve of the sample awaiting detection is greater than the reference curvature, it is determined that the sample awaiting detection has the prozone phenomenon, and if the calculated curvature of the reaction curve of the sample awaiting detection is equal to or less than the reference curvature, it is determined that the sample awaiting detection does not have the prozone phenomenon.

[0045] If it is determined that the sample to be detected does not have the prozone phenomenon ("No" in S200), the absorbance corresponding to a preset time point on the reaction curve of the sample to be detected is determined, and the antigen concentration of the sample to be detected corresponding to the absorbance of the sample to be detected according to the second calibration curve is determined. At this time, the determined antigen concentration of the sample to be detected can be directly determined as the antigen concentration of the sample to be detected.

[0046] In another embodiment, instead of using the second calibration curve, the antigen concentration of the sample to be detected can be calculated according to the Beer-Lambert law. If it is known that the absorbance of the reference sample with prozone phenomenon is A1 and the actual antigen concentration is M1, then the absorbance value of the sample to be detected is Ax, and the antigen concentration of the sample to be detected is Mx=M1×Ax÷A1 due to the direct proportionality between the absorbance and the actual antigen concentration.

[0047] If it is determined that the sample to be detected does not have the prozone phenomenon ("No" in S200), the antigen concentration of the sample to be detected corresponding to the calculated curvature of the reaction curve of the sample to be detected is within the interval OA in Figure 2, where the coordinates of O are (0,0) and the abscissa of point A corresponds to the upper limit of the linear range of CEA, and therefore the coordinates of point A are (100,0).

[0048] Conversely, if it is determined that the sample to be detected has the prozone phenomenon ("Yes" in S200), the antigen concentration of the sample to be detected corresponding to the curvature or length of the reaction curve of the sample to be detected is determined according to the first calibration curve shown in FIG. 2. At this time, there are two possibilities for the determined antigen concentration of the sample to be detected. One possibility is that the determined antigen concentration of the sample to be detected has only one value and is within the section AB, and the other possibility is that the determined antigen concentration of the sample to be detected has two values ​​and the antigen concentration determined by the two values ​​is within the section BC. The coordinates of point B are (492.5623, 0), and the coordinates of point C are (9000, 0). If the determined antigen concentration of the sample to be detected has one value, the determined antigen concentration of the sample to be detected is determined as the reference value of the antigen concentration of the sample to be detected.

[0049] When the determined antigen concentration of the sample to be detected has two values, the determined antigen concentration of the sample to be detected is verified, and the antigen concentration of the sample to be detected that is successfully verified is determined as the reference value of the antigen concentration of the sample to be detected.

[0050] In the verification, the lower of the determined antigen concentrations of the samples awaiting detection is verified, and if the verification is successful, the lower antigen concentration of the sample awaiting detection is determined as the reference value of the antigen concentration of the sample awaiting detection, and if the verification is unsuccessful, the higher antigen concentration of the sample awaiting detection is determined as the reference value of the antigen concentration of the sample awaiting detection.

[0051] The lower antigen concentration value of the sample to be detected is verified preferentially because the sample to be detected needs to be diluted in the verification process so that the sample to be detected is a sample free of the prozone phenomenon, and based on the lower antigen concentration of the sample to be detected, it can be diluted at a lower dilution rate until it is free of the prozone phenomenon.

[0052] Specifically, the step of verifying the lower of the determined antigen concentrations of the sample to be detected includes a step of diluting the sample to be detected so that the sample to be detected is a sample free of the prozone phenomenon (i.e., the concentration is diluted to the interval OA), and estimating the antigen concentration of the diluted sample to be detected based on the lower of the determined antigen concentrations of the sample to be detected.

[0053] Calculate the antigen concentration of the diluted sample to be detected. Based on the absorbance of the diluted sample to be detected, the second calibration curve is used to determine the antigen concentration of the diluted sample to be detected. Alternatively, based on the absorbance of the diluted sample to be detected, the antigen concentration of the diluted sample to be detected can be directly calculated using the Beer-Lambert law.

[0054] The estimated diluted antigen concentration of the sample to be detected is compared with the determined / calculated diluted antigen concentration of the sample to be detected, and if the estimated diluted antigen concentration of the sample to be detected matches the determined diluted antigen concentration of the sample to be detected, it indicates that the verification of the lower antigen concentration of the sample to be detected has been successful. Conversely, if the estimated diluted antigen concentration of the sample to be detected does not match the determined / calculated antigen concentration of the sample to be detected, it indicates that the verification of the lower antigen concentration of the sample to be detected has failed, and the higher antigen concentration of the sample to be detected is determined as the reference value of the antigen concentration of the sample to be detected.

[0055] For example, in Figure 2, when the curvature of the sample to be detected is equal to 2.7950m^-1, the corresponding antigen concentration of the sample to be detected has two values, 492.5623ng / ml and 9000ng / ml, respectively. Assuming that the antigen concentration is the lower of the two values, i.e., 492.5623ng / ml, and the liquid of the sample to be detected is diluted 6 times, the estimated antigen concentration of the diluted sample to be detected is about 82ng / ml, and if the concentration is diluted to less than 100ng / ml, the diluted sample to be detected does not have the prozone phenomenon. Based on the absorbance of the diluted sample to be detected, the second calibration curve or Lambert-Beer's law is used to obtain the antigen concentration of the diluted sample to be detected, and the obtained antigen concentration is compared with 82ng / ml. If the obtained antigen concentration is also 82ng / ml, the assumption that the antigen concentration is 492.5623ng / ml is correct, and the verification to the antigen concentration of 492.5623ng / ml is successful. If the obtained antigen concentration is not 82 ng / ml, the assumption that the antigen concentration is 492.5623 ng / ml is incorrect and the verification to an antigen concentration of 492.5623 ng / ml fails, thereby indicating that the antigen concentration of the sample to be detected is 9000 ng / ml.

[0056] 4 is a block diagram showing the configuration of a quantification device for detecting the prozone phenomenon in a sample to be detected according to an embodiment of the present invention. As shown in FIG. 4, the quantification device for detecting the prozone phenomenon in a sample to be detected according to an embodiment of the present invention includes a curve construction module, a confirmation module, a first quantification module, and a second quantification module. The curve construction module may be configured to construct a first calibration curve showing the relationship between the curvature of the reaction curve and the actual antigen concentration, or a first calibration curve showing the relationship between the length of the reaction curve and the actual antigen concentration, using the actual antigen concentration and the calculated curvature or length of the reaction curve of the change in absorbance over time of each of a plurality of reference samples including a sample with a prozone phenomenon and a sample without a prozone phenomenon. The curve construction module may further be configured to construct a second calibration curve showing the relationship between the absorbance and the actual antigen concentration, using the absorbance and the actual antigen concentration of each of the plurality of reference samples.

[0057] The determination module is configured to determine whether the sample to be detected has a prozone phenomenon based on the calculated curvature or length of the reaction curve of the sample to be detected. The first quantification module is configured to determine an antigen concentration of the sample to be detected corresponding to the curvature or length of the reaction curve of the sample to be detected according to the constructed first calibration curve when it is determined that the sample to be detected has a prozone phenomenon, determine the determined antigen concentration of the sample to be detected as a reference value of the antigen concentration of the sample to be detected when the determined antigen concentration of the sample to be detected is one value, and verify the determined antigen concentration of the sample to be detected as a reference value of the antigen concentration of the sample to be detected when the determined antigen concentration of the sample to be detected is two values. The second quantification module is configured to determine an antigen concentration of the sample to be detected corresponding to the absorbance of the sample to be detected according to the constructed second calibration curve when it is determined that the sample to be detected does not have a prozone phenomenon, and determine the determined antigen concentration of the sample to be detected as the antigen concentration of the sample to be detected.

[0058] Specifically, the first quantification module is configured to verify the lower of the determined antigen concentrations of the samples to be detected, and if the verification is successful, to determine the lower antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected, and if the verification is unsuccessful, to determine the higher antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected.

[0059] When verifying the lower value of the determined antigen concentrations of the samples to be detected, the first quantification module dilutes the sample to be detected so that the sample to be detected is a sample without the prozone phenomenon, and estimates the antigen concentration of the diluted sample to be detected based on the lower value of the antigen concentrations of the samples to be detected. The first quantification module determines the antigen concentration of the diluted sample to be detected using a second calibration curve based on the absorbance of the diluted sample to be detected. The first quantification module compares the estimated antigen concentration of the diluted sample to be detected with the determined antigen concentration of the diluted sample to be detected, and indicates that the verification of the lower antigen concentration of the sample to be detected is successful if the estimated antigen concentration of the diluted sample to be detected matches the determined antigen concentration of the diluted sample to be detected, and indicates that the verification of the lower antigen concentration of the sample to be detected is unsuccessful if the estimated antigen concentration of the diluted sample to be detected does not match the determined antigen concentration of the diluted sample to be detected.

[0060] The quantification device for detecting the prozone phenomenon in a sample to be detected according to an embodiment of the present invention further includes a calculation module, which is configured to calculate the curvature or length of the reaction curve of the absorbance change over time, to use an optical detection system in a reaction process to collect data of the transmitted light intensity change over time, convert the data of the transmitted light intensity change over time into a reaction curve of the absorbance change over time, determine an absorbance corresponding to a preset time point according to the reaction curve of the absorbance change over time, select a preset start time point and a preset end time point on the reaction curve of the absorbance change over time, and calculate the curvature or length of the reaction curve of the absorbance change over time between the preset start time point and the preset end time point.

[0061] According to an embodiment of the present invention, a qualitative prompt is given for samples to be detected that have prozone phenomenon, and a reference value for antigen concentration is given according to the length or curvature of the reaction curve, and especially for samples to be detected that have prozone phenomenon, the reference value for antigen concentration can provide a reference for the actual antigen concentration that is excessive.

[0062] The various embodiments of the present invention are not intended to be an exhaustive enumeration of all possible combinations, but are intended to describe representative aspects of the present invention, and the contents described in each embodiment may be applied independently or in combination of two or more.

[0063] The description given in the above exemplary embodiments is only for illustrating the technical solutions of the present invention, and is not intended to be exhaustive, and is not intended to limit the present invention to the described forms. Obviously, those skilled in the art can make many modifications and variations according to the above teachings. The exemplary embodiments are selected and described to explain the particular principles of the present invention and its practical applications, so as to facilitate other skilled in the art to understand and actually utilize the various exemplary embodiments of the present invention and their various alternatives and modifications. The protection scope of the present invention is intended to be defined by the appended claims and their equivalents.

Claims

1. 1. A quantification method for detecting prozone phenomena in a sample to be detected, comprising: A step of constructing a first calibration curve showing the relationship between the curvature of the reaction curve and the actual antigen concentration, or a first calibration curve showing the relationship between the length of the reaction curve and the actual antigen concentration, using the actual antigen concentration and the curvature or length of the reaction curve of the calculated change in absorbance over time of each of a plurality of reference samples including a sample with a prozone phenomenon and a sample without a prozone phenomenon; determining whether the sample to be detected has a prozone phenomenon based on the calculated curvature or length of the response curve of the sample to be detected; A quantification method for detecting the prozone phenomenon in a sample to be detected, comprising the steps of: when it is determined that the sample to be detected has the prozone phenomenon, determining an antigen concentration of the sample to be detected corresponding to a curvature or length of the reaction curve of the sample to be detected according to the constructed first calibration curve; when the determined antigen concentration of the sample to be detected is one value, determining the determined antigen concentration of the sample to be detected as a reference value of the antigen concentration of the sample to be detected; and when the determined antigen concentration of the sample to be detected is two values, verifying the determined antigen concentration of the sample to be detected and determining the antigen concentration of the sample to be detected that has been successfully verified as the reference value of the antigen concentration of the sample to be detected.

2. constructing a second calibration curve showing the relationship between absorbance and actual antigen concentration using the absorbance and actual antigen concentration of each of the plurality of reference samples; 2. The quantification method for detecting the prozone phenomenon in a sample to be detected as described in claim 1, further comprising a step of: when it is determined that the sample to be detected does not have the prozone phenomenon, determining an antigen concentration of the sample to be detected corresponding to the absorbance of the sample to be detected according to the constructed second calibration curve, and determining the determined antigen concentration of the sample to be detected as the antigen concentration of the sample to be detected.

3. When the determined antigen concentration of the sample to be detected is two values, the step of verifying the determined antigen concentration of the sample to be detected and determining the antigen concentration of the sample to be detected that has been successfully verified as the reference value of the antigen concentration of the sample to be detected includes: A quantification method for detecting the prozone phenomenon in a sample to be detected as described in claim 2, characterized in that it includes a step of verifying the lower of the determined antigen concentrations of the sample to be detected, and if the verification is successful, determining the lower antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected, and if the verification is unsuccessful, determining the higher antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected.

4. The step of verifying the lower value of the determined antigen concentrations of the samples to be detected includes: A step of diluting the sample to be detected so that the sample to be detected is a sample free of the prozone phenomenon, and estimating the antigen concentration of the diluted sample to be detected based on the lower value of the antigen concentrations of the samples to be detected; determining an antigen concentration of the diluted sample to be detected using a second calibration curve based on the absorbance of the diluted sample to be detected; 4. The method for quantifying prozone phenomenon in a sample to be detected according to claim 3, further comprising the steps of: comparing the estimated diluted antigen concentration of the sample to be detected with the determined diluted antigen concentration of the sample to be detected; and indicating that verification of the lower antigen concentration of the sample to be detected has been successful if the estimated diluted antigen concentration of the sample to be detected matches the determined diluted antigen concentration of the sample to be detected; and indicating that verification of the lower antigen concentration of the sample to be detected has failed if the estimated diluted antigen concentration of the sample to be detected does not match the determined diluted antigen concentration of the sample to be detected.

5. Calculating the curvature or length of the response curve of the change in absorbance over time includes: Using an optical detection system to collect data of the change in transmitted light intensity over time during the reaction process, converting the data of the change in transmitted light intensity over time into a response curve of the change in absorbance over time, and determining the absorbance corresponding to a preset time point according to the response curve of the change in absorbance over time; A quantification method for detecting the prozone phenomenon in a sample to be detected as described in claim 2, characterized in that it includes a step of selecting a preset start point and a preset end point on a reaction curve of the change in absorbance over time, and calculating the curvature or length of the reaction curve of the change in absorbance over time between the preset start point and the preset end point.

6. 1. A quantification device for detecting the prozone phenomenon in a sample to be detected, comprising: a curve construction module configured to construct a first calibration curve showing a relationship between the curvature of the reaction curve and the actual antigen concentration, or a first calibration curve showing a relationship between the length of the reaction curve and the actual antigen concentration, using the actual antigen concentration and the curvature or length of the reaction curve of the calculated change in absorbance over time of each of a plurality of reference samples including a sample with a prozone phenomenon and a sample without a prozone phenomenon; a determination module configured to determine whether the sample to be detected has a prozone phenomenon based on the calculated curvature or length of the response curve of the sample to be detected; a first quantification module configured to, when it is determined that the sample to be detected has a prozone phenomenon, determine an antigen concentration of the sample to be detected corresponding to a curvature or length of a reaction curve of the sample to be detected according to a constructed first calibration curve, and, when the determined antigen concentration of the sample to be detected is one value, determine the determined antigen concentration of the sample to be detected as a reference value of the antigen concentration of the sample to be detected, and, when the determined antigen concentration of the sample to be detected is two values, verify the determined antigen concentration of the sample to be detected, and determine the antigen concentration of the sample to be detected that has been successfully verified as the reference value of the antigen concentration of the sample to be detected.

7. The curve construction module is further configured to construct a second calibration curve showing a relationship between the absorbance and the actual antigen concentration using the absorbance and the actual antigen concentration of each of the plurality of reference samples; The quantification device for detecting the prozone phenomenon in a sample to be detected as described in claim 6, further comprising a second quantification module, which is configured to determine an antigen concentration of the sample to be detected corresponding to the absorbance of the sample to be detected according to the constructed second calibration curve when it is determined that the sample to be detected does not have the prozone phenomenon, and to determine the determined antigen concentration of the sample to be detected as the antigen concentration of the sample to be detected.

8. The quantification device for detecting the prozone phenomenon in a sample to be detected as described in claim 7, characterized in that the first quantification module is configured to verify the lower value of the determined antigen concentrations of the sample to be detected, and if the verification is successful, to determine the lower antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected, and if the verification is unsuccessful, to determine the higher antigen concentration of the sample to be detected as the reference value of the antigen concentration of the sample to be detected.

9. The first quantification module: When verifying the lower value of the determined antigen concentrations of the samples to be detected, dilute the samples to be detected so that the samples to be detected are free of the prozone phenomenon, and estimate the antigen concentration of the diluted samples to be detected based on the lower value of the antigen concentrations of the samples to be detected; determining an antigen concentration of the diluted sample to be detected using a second calibration curve based on the absorbance of the diluted sample to be detected; The quantification device for detecting the prozone phenomenon in a sample to be detected as described in claim 8, which is configured to compare the estimated antigen concentration of the diluted sample to be detected with the determined antigen concentration of the diluted sample to be detected, and if the estimated antigen concentration of the diluted sample to be detected matches the determined antigen concentration of the diluted sample to be detected, indicate that the verification of the lower antigen concentration of the sample to be detected has been successful, and if the estimated antigen concentration of the diluted sample to be detected does not match the determined antigen concentration of the diluted sample to be detected, indicate that the verification of the lower antigen concentration of the sample to be detected has failed.

10. The apparatus includes a calculation module for calculating a curvature or length of a response curve of a change in absorbance over time, the calculation module comprising: Using an optical detection system in the reaction process to collect data on the change in transmitted light intensity over time, converting the data on the change in transmitted light intensity over time into a response curve of the change in absorbance over time, and determining the absorbance corresponding to a preset time point according to the response curve of the change in absorbance over time; A quantification device for detecting the prozone phenomenon in a sample to be detected, as described in claim 7, characterized in that it is configured to select a preset start point and a preset end point on a reaction curve of the change in absorbance over time, and calculate the curvature or length of the reaction curve of the change in absorbance over time between the preset start point and the preset end point.

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