Method and device for detecting viral antigen of C type hepatitis
A technology for hepatitis C virus and antigen, which is applied in the field of hepatitis C virus detection, can solve the problems of difficult vaccine, difficult hepatitis C detection, and no protective effect, and achieves high specificity, good application prospects, and low false negatives. rate and false positive rate
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Embodiment 1
[0021] Detecting hepatitis C virus antigen method, described method comprises the following steps: (1) antibody coating: I-hepatitis C virus-IgG; (2) the antigen in the sample is combined with coating antibody: I-hepatitis C virus -IgG-hepatitis C virus; (3) the hepatitis C virus antibody of enzyme label is combined with antigen: I-hepatitis C virus-IgG-hepatitis C virus-IgG-HRP; (4) add peroxidase ( HRP) substrate solution color development; (5) with 2mol / L of H 2 SO 4 Terminate the reaction; (6) microplate reader detection.
Embodiment 2
[0023] The preparation method of hepatitis C virus antigen detection kit:
[0024] (1) Antigen for immunization: The antigen used in the preparation of the antibody in the detection kit is a highly purified Hepatitis C virus recombinant gene containing 10 highly conserved regions of hepatitis C virus expressed by Escherichia coli using molecular biology techniques. Hepatitis virus multi-epitope complex antigen. The antigen has good immunogenicity and can induce animals to produce high-titer specific antibody responses against the hepatitis C virus. Due to the highly conserved nature of the immunogen, the prepared antibody can recognize different types of hepatitis C virus.
[0025] (2) Preparation of specific antibody: use clean grade Japanese white rabbits, weighing 1.8-2.0Kg, inject 1 mg / ml compound antigen and the same amount of incomplete Freund's adjuvant, fully emulsify and mix, and inject into muscles at multiple points. The immunization procedure was 3 times in 0, 1,...
Embodiment 3
[0031] Sensitivity and specificity test of hepatitis C virus antigen detection kit: based on the review of hepatitis C virus-RNA detection. Detect the negative and positive samples of hepatitis C antigen (HCAg) with the kit described in Example 2, extract hepatitis C virus-RNA with a small amount of virus RNA / DNA extraction kit, and then use RT-PCR detection kit to carry out HCV-RNA detection to determine its sensitivity (true positive rate), specificity (true negative rate), false positive rate, false negative rate, crude consistency, adjusted consistency, Youden index, positive predictive value and Negative predictive value. A total of 400 samples were compared, and the results are shown in Table 1.
[0032] Table 1.
[0033] RT-PCR method detection results
[0034] Kit HCAg test results Total
[0035] Positive Negative
[0036] Positive 34 3 37
[0037] Female 5 258 363
[0038] Total 39 361 400
[0039] in:
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