Efficient hepatitis C virus core antigen enzyme-linked immunosorbent assay detection kit
A hepatitis C virus, enzyme-linked immunoassay detection technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of insufficient specificity and low sensitivity, and achieve improved analytical sensitivity, good solubility and dispersion, and improved stability. sexual effect
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Embodiment 1
[0037] The composition and preparation method of the kit
[0038] An enzyme-linked immunosorbent assay kit for hepatitis C virus core antigen includes:
[0039] 1 96-well anti-HCV-cAg monoclonal antibody coated plate, anti-HCV-cAg enzyme conjugate, 5 bottles of standard, one copy of negative and positive control, HCV-cAg sample diluent, substrate solution A, substrate 1 bottle each of liquid B, stop liquid, and washing liquid (20×concentrated liquid), an instruction sheet, and 4 stickers.
[0040] 1. Coated with anti-HCV-cAg monoclonal antibodies Mab1 and Mab2 to form a reaction plate. The ELISA plate is a milky white opaque polystyrene 96-well ELISA plate.
[0041] Coating method of antibody on 96-well ELISA plate: Dilute the purified Mab1 and Mab2 with pH5.0 tartaric acid buffer to 4μg / mL and 4μg / mL respectively, and add 100μL to each well of the detection plate. Place it at 2-8°C for 12 hours, discard the coating solution, wash it 5-8 times with the washing solution, and then gent...
Embodiment 2
[0053] Application of the kit and testing procedures
[0054] (1) Equilibrium: equilibrate the sample and the reagents in the test kit at 18-25℃ for 30 minutes;
[0055] (2) Preparation: Dilute the concentrated washing solution in the kit 20 times with deionized water, measure 300 mL, and prepare the washing solution of the concentration required for the work;
[0056] (3) Sample diluent: add sample diluent and sample at a ratio of 1:2 to each well on the pretreatment plate, incubate with shaking at 45-60°C for 60 minutes, and set aside;
[0057] (4) Sample addition: Take a 96-well ELISA plate, preset 3 wells each for blank, negative, and positive controls. Add 100μl of sample diluent to each well of the blank control, and add 50μl of sample diluent to each well of the remaining wells, and then negative and positive controls Add 50μl of the corresponding control serum to each well, and test the remaining wells. Add 50μl of each sample to be tested after the above pretreatment accordin...
Embodiment 3
[0065] The precision, specificity and sensitivity of this kit
[0066] 1) Precision test: using PCR and EIA to determine the negative and positive sera for 20 points each, using 20 negative sera to test, no case is positive; using 20 positive sera to test, no case is negative, which meets the product standard.
[0067] 2) Specificity test: The results of the cross-reaction verification test show that anti-HAV, HBsAg, anti-TP, anti-TOX, HSV, CMV, EBV, positive samples and samples from patients with primary liver cancer do not affect the test results.
[0068] 27 cases of RF positive samples, 25 cases of HAMA positive samples, 18 cases of ANA positive samples (the above samples are negative for anti-HCV and HCV) were tested for HCV-cAg using the present invention. The number of false positives for the three types of samples was RF false positives. 0, HAMA false positive is 1, ANA false positive is 1, the specificity is 100%, 96%, 94.44%, respectively.
[0069] 3) Sensitivity test: the s...
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