A fluorescent immunoassay analyzer quality control card and its preparation method and application

Abstract

The present invention belongs to the technical field of quality control cards, and specifically relates to a quality control card for a fluorescent immunoassay analyzer, a preparation method thereof, and an application thereof. The present invention provides a quality control card for a fluorescent immunoassay analyzer, comprising a quality control reaction membrane, on which a quality control product and a fluorescently labeled antibody are carried. The present invention uses a reaction membrane carrying a quality control product and a fluorescently labeled antibody as a raw material to prepare a quality control card for a fluorescent immunoassay analyzer, wherein the quality control product and the fluorescently labeled antibody undergo an antigen-antibody reaction, so that the fluorescent substance is stable, the structure is reasonable, and the card can be stably stored, thereby extending the service life of the quality control card, ensuring calibration accuracy, and saving costs.

Description

Technical Field

[0001] The invention belongs to the technical field of quality control cards, and in particular relates to a quality control card for a fluorescent immunoassay analyzer, a preparation method thereof and applications thereof. Background Art

[0002] Immunochromatography is a new type of rapid immunoassay technology with the characteristics of rapidity, accuracy, and quantification. It is widely used in the identification and detection of markers in the fields of medical diagnosis, environmental monitoring, and food safety. Due to its advantages such as high sensitivity, good stability, and easy quantification, the immunofluorescence method has gradually replaced the traditional colloidal gold method and is widely used. Fluorescence immunochromatography detection requires test plates and fluorescence quantitative analyzers. Immunofluorescence quantitative analyzers have been widely used, but calibration and traceability still have problems such as lack of standard fluorescence intensity cards and inconsistent calibration methods, which seriously affect the accuracy and repeatability of measurements.

[0003] At present, cards marked with fluorescent molecules (or fluorescent quantum dots) or LED light sources with specific wavelengths are usually used as calibration cards. Since the fluorescence of fluorescent molecules or quantum dots is greatly affected by the surrounding environment, it is difficult to ensure stability, which makes it difficult to formulate standards and conduct long-term quality control. LED lights themselves have differences in luminous intensity and color, and standard cards made with LED lights or light strips have poor consistency. Summary of the invention

[0004] The purpose of the present invention is to solve the deficiencies in the prior art and provide a quality control card for a fluorescent immunoassay analyzer and a preparation method thereof to improve stability and accuracy.

[0005] The invention provides a quality control card for a fluorescent immunoassay analyzer. The quality control card comprises a quality control reaction membrane on which a quality control product and a fluorescently labeled antibody are carried.

[0006] Preferably, the quality control reaction membrane comprises a quality control NC membrane.

[0007] Preferably, the fluorescent immunoassay analyzer quality control card further comprises a reagent shell for assembling the quality control reaction membrane.

[0008] Preferably, the fluorescently labeled antibody contains an antibody that specifically binds to the quality control product.

[0009] The present invention also provides a method for preparing the fluorescent immunoassay analyzer quality control card described in the above technical solution, comprising the following steps:

[0010] After mixing and incubating a solution containing a quality control product with a reaction membrane containing a fluorescently labeled antibody, taking out the reaction membrane and drying it to obtain the quality control reaction membrane;

[0011] The quality control reaction membrane is placed in a reagent shell to obtain the fluorescence immunoassay analyzer quality control card.

[0012] Preferably, the method of mixing the solution containing the quality control product with the reaction membrane comprises: adding the solution containing the quality control product into a fluorescent immunoassay plate, wherein the fluorescent immunoassay plate contains the reaction membrane.

[0013] Preferably, the concentration of the quality control product in the solution containing the quality control product is 4-6 ng / mL.

[0014] Preferably, the incubation time is 15 minutes; the drying temperature is 45° C. and the drying time is 3 hours.

[0015] Preferably, after the quality control reaction membrane is loaded into the reagent shell, the method further comprises: performing QR code spraying.

[0016] The present invention also provides the use of the fluorescence immunoassay analyzer quality control card described in the above technical solution in the preparation of an electronic quality control card assembly for a fluorescence immunoassay analyzer.

[0017] The present invention provides a fluorescent immunoassay instrument quality control card, comprising a quality control reaction membrane, on which a quality control product and a fluorescently labeled antibody are carried. The present invention uses the reaction membrane carrying the quality control product and the fluorescently labeled antibody as a raw material to prepare a fluorescent immunoassay instrument quality control card, wherein the quality control product and the fluorescently labeled antibody undergo antigen-antibody reaction, so that the fluorescent substance is stable, the structure is reasonable, and the card can be stably stored, thereby extending the service life of the quality control card, ensuring the accuracy of calibration, and saving costs.

[0018] Furthermore, the fluorescent immunoassay analyzer quality control card prepared using the quality control product as a raw material of the present invention can quantitatively determine the content of the quality control product and serve as a positive control for the sample to be tested. DETAILED DESCRIPTION

[0019] The invention provides a quality control card for a fluorescent immunoassay analyzer, comprising a quality control reaction membrane on which a quality control product and a fluorescently labeled antibody are carried.

[0020] The quality control reaction membrane of the present invention preferably includes a quality control NC membrane. The present invention has no strict requirements on the type of the quality control product, and the quality control products conventionally used in fluorescent immunoassay can be selected, including but not limited to any one of brain natriuretic peptide (BNP) quality control products, troponin I (TnⅠ) quality control products and D-dimer (D-Dimer) quality control products. The present invention uses troponin I quality control products as an example for explanation in the specific implementation process, but it cannot be understood as the entire protection scope of the present invention. The present invention has no strict requirements on the source of the quality control product, and it can be purchased conventionally. The troponin I (TnⅠ) quality control product used in the specific implementation process of the present invention is preferably purchased from Hebei Twente Biotechnology Development Co., Ltd., with batch number FC220101.

[0021] The fluorescently labeled antibody of the present invention preferably contains an antibody that specifically binds to the quality control product. The present invention has no strict requirements on the source of the fluorescently labeled antibody, and conventional purchase is sufficient. The present invention uses the fluorescently labeled antibody as a raw material, which can bind to the quality control product by antigen-antibody, so that the fluorescent substance exists stably.

[0022] In the present invention, the fluorescent immunoassay analyzer quality control card preferably also includes a reagent shell for assembling the quality control reaction membrane; the reagent shell is preferably sprayed with a QR code containing quality control standard information. The present invention sprays the QR code on the reagent shell to distinguish the batch information of different quality control cards, and after quality control analysis, distinguishes the fluorescence intensity of different batches of quality control cards.

[0023] The present invention uses a reaction membrane carrying a quality control product as a raw material to prepare a fluorescent immunoassay quality control card, so that the luminescent chemical substance is stable, the structure is reasonable, and it can be stably stored, and the repeatability, stability, and accuracy of the equipment can be quantitatively judged, and the cost can be saved.

[0024] The present invention also provides a method for preparing the above-mentioned fluorescent immunoassay instrument quality control card, comprising the following steps:

[0025] After mixing and incubating the solution containing the quality control product with the antibody reaction membrane containing the fluorescent marker, taking out the reaction membrane, and drying it to obtain the quality control reaction membrane;

[0026] The quality control reaction membrane is placed in a reagent shell, and two-dimensional code is sprayed to obtain the fluorescence immunoassay analyzer quality control card.

[0027] The present invention mixes and incubates a solution containing a quality control product with a reaction membrane. The mixing method of the present invention preferably includes: adding a solution containing a quality control product to a fluorescent immunoassay plate, wherein the fluorescent immunoassay plate contains a reaction membrane, and the reaction membrane contains a fluorescently labeled antibody; the fluorescent immunoassay plate is preferably taken from a fluorescent detection kit. The incubation time of the present invention is preferably 15 minutes. The present invention preferably selects a fluorescent immunoassay plate for use with the type of quality control product. Specifically, when the quality control product is a troponin I quality control product, the fluorescent immunoassay plate is preferably taken from a troponin I detection kit; when the quality control product is a brain natriuretic peptide quality control product, the fluorescent immunoassay plate is preferably taken from a brain natriuretic peptide detection kit; when the quality control product is a D-dimer quality control product, the fluorescent immunoassay plate is preferably taken from a D-dimer detection kit. The present invention has no strict requirements on the source of the fluorescent immunoassay kit, and conventional purchase is sufficient. The present invention is described by taking troponin I quality control products and troponin I detection kit as examples in the specific implementation process, but it cannot be understood as the entire protection scope of the present invention. The troponin I detection kit (fluorescence immunoassay) is preferably purchased from Hebei Twente Biotechnology Development Co., Ltd., with batch number T211101. The detection plate of the troponin I detection kit (fluorescence immunoassay) of the present invention contains the reaction membrane, and the reaction membrane is coated with fluorescently labeled antibodies. By adding the quality control product to the detection plate and incubating for 15 minutes, the quality control product and the fluorescently labeled antibody can be closely combined to stabilize the fluorescent structure.

[0028] After the incubation is completed, the present invention removes the reaction membrane and obtains the quality control reaction membrane after drying. The drying time of the present invention is preferably 3 hours, and the drying temperature is preferably 45°C; the drying is preferably carried out in an oven. The present invention has no strict requirements on the method of removing the reaction membrane, and conventional operations are sufficient. The present invention can make the fluorescent conjugate more stable by drying.

[0029] After obtaining the quality control reaction membrane, the present invention loads the quality control reaction membrane into a reagent shell, performs two-dimensional code spraying, and obtains the fluorescent immunoassay instrument quality control card. The present invention preferably uses welding to load the quality control reaction membrane into the reagent shell, and further preferably uses an electric welder. The two-dimensional code of the present invention preferably contains quality control standard information, that is, the batch information of the fluorescent immunoassay instrument quality control card. The present invention has no strict requirements on the method of spraying the two-dimensional code, and conventional operation is sufficient.

[0030] The present invention adds a quality control product to a test plate for incubation, and uses the test plate to which the quality control product has been added and reacted completely, takes out its reaction membrane, dries it, and reassembles it into a quality control card, which can stabilize the fluorescent substance, has a reasonable structure, can be stably stored, has good repeatability, high accuracy, and saves costs. Compared with directly using the test plate of the test kit (fluorescent immunoassay) for calibration, it can save time and cost.

[0031] The present invention also provides the use of the fluorescent immunoassay analyzer quality control card described in the above technical solution in the preparation of an electronic quality control card assembly for a fluorescent immunoassay analyzer. The fluorescent immunoassay analyzer quality control card of the present invention can be used to prepare an electronic quality control card assembly for a fluorescent immunoassay analyzer, perform fluorescent immunoassay, and has good stability and high calibration accuracy.

[0032] In order to further illustrate the present invention, a fluorescent immunoassay analyzer quality control card, a preparation method and an application thereof provided by the present invention are described in detail below in conjunction with embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0033] Example 1

[0034] The preparation method of the quality control card of the fluorescent immunoassay analyzer consists of the following steps:

[0035] 1. Place the troponin I detection kit (fluorescent immunoassay, batch number T211101) and the solution containing troponin I quality control product (batch number FC220101) of Hebei Twente Biotechnology Development Co., Ltd. at room temperature for 30 minutes. The concentration of troponin I quality control product in the solution containing troponin I quality control product is 5±1ng / mL;

[0036] 2. Add 200 μL of the solution containing troponin I quality control product to the RM membrane of the troponin I detection kit (i.e., the NC membrane carrying the fluorescent-labeled antibody) and react for 15 minutes;

[0037] 3. Take out the RM membrane from the reagent kit after the reaction is complete and put it into a 45℃ oven to dry for 3 hours;

[0038] 4. Put the dried RM membrane into the new reagent shell and weld it using a welding machine;

[0039] 5. Spray the QR code on the reagent shell. The QR code is the batch information of the quality control card.

[0040] Comparative Example 1

[0041] Same as Example 1, except that the concentration of the troponin I control substance in the solution containing the troponin I control substance in step 1 is 1±0.2 ng / mL.

[0042] Comparative Example 2

[0043] Same as Example 1, except that step 3 is to put the mixture into a 45°C oven and dry it for 2 h.

[0044] Comparative Example 3

[0045] Same as Example 1, except that step 3 is to put the mixture into a 45°C oven and dry it for 3.5 hours.

[0046] Comparative Example 4

[0047] Same as Example 1, except that step 3 is to put the mixture into a 40° C. oven and dry it for 3 h.

[0048] Comparative Example 5

[0049] Same as Example 1, except that step 3 is to put the mixture into a 50° C. oven and dry it for 3 h.

[0050] Test Example 1

[0051] Accuracy test

[0052] Take 5 quality control cards from the same batch as Example 1 and Comparative Examples 1 to 5 as test samples, respectively, insert the quality control cards into a fluorescent immunoassay analyzer (model: T-Meter I), and the analyzer recognizes the QR code on the quality control card, which is the parameter data (batch information) of the quality control card. The analyzer automatically scans the fluorescence line on the quality control card and automatically reads the fluorescence values ​​line1 and line2. Each test sample is tested 3 times, and the relative deviation (%) is calculated to evaluate the accuracy of the quality control card detection. The results are shown in Tables 1 to 6.

[0053] Table 1 Accuracy test results of the quality control card obtained in Example 1

[0054]

[0055] Table 2 Accuracy test results of the quality control card obtained in comparative example 1

[0056]

[0057]

[0058] Table 3 Accuracy test results of the quality control card obtained in comparative example 2

[0059]

[0060] Table 4 Accuracy test results of the quality control card obtained in Example 3

[0061]

[0062]

[0063] Table 5 Accuracy test results of the quality control card obtained in Example 4

[0064]

[0065] Table 6 Accuracy test results of the quality control card obtained in Example 5

[0066]

[0067] According to Tables 1 to 6, it can be seen that the relative deviations of the quality control cards prepared in Example 1 are all within 4%, the relative deviation of the quality control cards prepared in Comparative Example 1 is 5.54% at most, the relative deviation of the quality control cards prepared in Comparative Example 2 is 5.85% at most, the relative deviation of the quality control cards prepared in Comparative Example 3 is 5.25% at most, the relative deviation of the quality control cards prepared in Comparative Example 4 is 5.83% at most, and the relative deviation of the quality control cards prepared in Comparative Example 5 is 5.48% at most. The quality control cards prepared by the present invention have good accuracy.

[0068] Test Example 2

[0069] Stability testing

[0070] Ten quality control cards from the same batch as Example 1 and Comparative Examples 1 to 5 were taken as test samples, and the fluorescence values ​​were continuously measured 10 times on the 1st day, 14th day, 1 month, 2 months, 3 months, 5 months, 6 months, 8 months, 10 months, and 12 months of preparation. The coefficient of variation CV of the 10 fluorescence values ​​was calculated according to the following formula to evaluate the stability of the quality control card detection. The results are shown in Tables 7 to 12.

[0071]

[0072] The average value of the measurement results

[0073] S: Standard deviation

[0074] Table 7 Stability test results of the quality control card obtained in Example 1

[0075]

[0076]

[0077] Table 8 Stability test results of the quality control card obtained in comparative example 1

[0078]

[0079]

[0080] Table 9 Stability test results of the quality control card obtained in comparative example 2

[0081]

[0082]

[0083] Table 10 Stability test results of the quality control card obtained in Example 3

[0084]

[0085] Table 11 Stability test results of the quality control card obtained in Example 4

[0086]

[0087]

[0088] Table 12 Stability test results of the quality control card obtained in Example 5

[0089]

[0090]

[0091] According to Tables 7 to 12, it can be seen that the coefficient of variation CV of the quality control card prepared in Example 1 is within 8% for 10 consecutive measurements within 12 months, the maximum coefficient of variation CV of the quality control card prepared in Comparative Example 1 is 10.34% for 10 consecutive measurements within 12 months, the maximum coefficient of variation CV of the quality control card prepared in Comparative Example 2 is 10.56% for 10 consecutive measurements within 12 months, the maximum coefficient of variation CV of the quality control card prepared in Comparative Example 3 is 10.18% for 10 consecutive measurements within 12 months, the maximum coefficient of variation CV of the quality control card prepared in Comparative Example 4 is 11.11% for 10 consecutive measurements within 12 months, and the maximum coefficient of variation CV of the quality control card prepared in Comparative Example 5 is 10.60% for 10 consecutive measurements within 12 months. The quality control card prepared by the present invention has good stability.

[0092] Test Example 3

[0093] Stability comparison

[0094] At 25°C, the quality control card obtained in Example 1 (denoted as TWT) and the fluorescent microsphere quality control card (210104) of Changsha Meiniu Biotechnology Co., Ltd. were stored at relative humidity of 20%, 30%, 40%, 50%, 60%, 70% and 80%, respectively. After storage for 7 days, 10 samples of Example 1 and the fluorescent microsphere quality control card were taken as test samples, and the fluorescence values ​​were measured 3 times. The coefficient of variation CV of the fluorescence value was calculated according to the formula of Test Example 2. The results are shown in Table 17.

[0095] Table 17 Stability test results of different quality control cards at different storage humidity

[0096]

[0097] It can be seen from Table 17 that the quality control card prepared by the present invention tolerates a wider humidity range, is easier to store, and has better stability.

[0098] It can be seen from the above content that the quality control card prepared by the present invention has good stability and high accuracy.

[0099] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing a quality control card for a fluorescent immunoassay analyzer, characterized in that: The steps include: After mixing and incubating a solution containing a quality control product with a reaction membrane containing a fluorescently labeled antibody, taking out the reaction membrane and drying it to obtain a quality control reaction membrane; the fluorescently labeled antibody contains an antibody that specifically binds to the quality control product; The quality control reaction membrane is placed in a reagent shell to obtain a fluorescence immunoassay analyzer quality control card; The concentration of the quality control product in the solution containing the quality control product is 4-6 ng / mL; The incubation time is 15 min; The drying temperature is 45° C. and the drying time is 3 hours.

2. The preparation method according to claim 1, characterized in that: The quality control reaction membrane includes a quality control NC membrane.

3. The preparation method according to claim 1, characterized in that: The method of mixing the solution containing the quality control product with the reaction membrane includes: adding the solution containing the quality control product into a fluorescent immunoassay plate, wherein the fluorescent immunoassay plate contains the reaction membrane.

4. The preparation method according to any one of claims 1 to 3, characterized in that: After the quality control reaction membrane is loaded into the reagent shell, the method further includes: performing QR code spraying.

5. Use of the quality control card for a fluorescent immunoassay analyzer obtained by the preparation method according to any one of claims 1 to 4 in the preparation of an electronic quality control card assembly for a fluorescent immunoassay analyzer.

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