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A highly efficient hepatitis C virus core antigen ELISA kit

A hepatitis C virus, enzyme-linked immunosorbent assay technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of low sensitivity and low specificity, and achieve improved analytical sensitivity, good solubility and dispersion, excellent Effects of washing and foaming

Active Publication Date: 2016-04-06
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problems of the existing hepatitis C antigen ELISA kits such as low sensitivity and insufficient specificity, the present invention provides an efficient hepatitis C virus core antigen ELISA kit

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The composition and preparation method of the kit

[0038] A hepatitis C virus core antigen ELISA kit comprises:

[0039] 1 piece of 96-well anti-HCV-cAg monoclonal antibody coated plate, anti-HCV-cAg enzyme conjugate, 5 bottles of standard product, one copy of negative control and one positive control, HCV-cAg sample diluent, substrate solution A, substrate 1 bottle each of Solution B, Stop Solution, and Washing Solution (20×concentrated solution), 1 copy of instructions, and 4 sheets of self-adhesive paper.

[0040] 1. Coat the anti-HCV-cAg monoclonal antibodies Mab1 and Mab2 to form a reaction plate, and choose a milky white opaque polystyrene 96-well microtiter plate.

[0041]Coating method of antibody on 96-well ELISA plate: Dilute the purified Mab1 and Mab2 to 4 μg / mL and 4 μg / mL respectively with pH 5.0 tartrate buffer, add 100 μL to each well of the assay plate, Place at 2-8°C for 12 hours, discard the coating solution, wash 5-8 times with washing solution, an...

Embodiment 2

[0053] Kit application and detection procedures

[0054] (1) Equilibration: equilibrate the sample and the reagents in the test kit at 18-25°C for 30 minutes;

[0055] (2) Dosing: dilute the concentrated washing liquid in the kit with deionized water 20 times, measure 300mL, and make the washing liquid with the concentration required for the work, and prepare for the following plate washing;

[0056] (3) Sample diluent: add sample diluent and sample to each well on the pretreatment plate at a ratio of 1:2, shake and incubate at 45-60°C for 60 minutes, and set aside;

[0057] (4) Adding samples: Take a 96-well ELISA plate, pre-set 3 holes each for blank, negative, and positive controls, add 100 μl of sample diluent to each well of the blank control, add 50 μl of sample diluent to each well of the remaining wells, and then add negative and positive controls. Add 50 μl of the corresponding control serum to each well, and add 50 μl of the samples to be tested in the remaining wel...

Embodiment 3

[0065] Detection of precision, specificity and sensitivity of this kit

[0066] 1) Precision test: use PCR and EIA to determine 20 points each for negative and positive sera, test with 20 negative sera, none of them are positive; test with 20 positive sera, none of them are negative, which meets the product standard.

[0067] 2) Specificity test: The results of the cross-reaction verification test showed that anti-HAV, HBsAg, anti-TP, anti-TOX, HSV, CMV, EBV, positive samples and samples from patients with primary liver cancer did not affect the detection results.

[0068] For 27 cases of RF positive samples, 25 cases of HAMA positive samples, and 18 cases of ANA positive samples (the above samples were both negative for HCV and HCV), respectively, the present invention was used to detect HCV-cAg, and 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, and the specificity is 100%, 96%, 94.44...

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PUM

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Abstract

The invention discloses an efficient hepatitis C virus (HCV) core antigen enzyme-linked immunosorbent assay (ELISA) detection kit which mainly comprises a microlon ELISA plate, an anti-HCV-cAg enzyme conjugate, an HCV-cAg standard substance gradient solution, a positive serum control indication sample, a negative serum control indication sample, an HCV-cAg sample diluent, a substrate solution A, a substrate solution B, a stop buffer and 20* concentrated washing liquid. The kit provided by the invention is high in sensitivity, good in precision and strong in specificity.

Description

technical field [0001] The invention relates to a virus antigen detection kit, in particular to an efficient hepatitis C virus core antigen (HCV-coreAg) ELISA detection kit. Background technique [0002] Hepatitis C virus, referred to as hepatitis C, hepatitis C, is a kind of viral hepatitis caused by hepatitis C virus (HepatitisCvirus, HCV) infection, hepatitis C is a global epidemic, can lead to chronic inflammation and necrosis of the liver and fibrous Some patients may develop liver cirrhosis or even hepatocellular carcinoma (HCC). Some data show that the mortality rate related to HCV infection (liver failure and death caused by hepatocellular carcinoma) will continue to increase in the next 20 years, which is extremely harmful to the health and life of patients. Currently, chronic hepatitis C has definite curative effects except interferon , there is no other effective treatment, and it is difficult to make a breakthrough in the development of vaccines in a short perio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/576
CPCG01N33/56983G01N33/5767
Inventor 谭柏清罗维晓甘宜梧李静赵新朱玉娥肖慧
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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