Kit for combined detection of hepatitis C virus antigen and antibody through chemiluminescence

A chemiluminescence and combined detection technology, applied in measurement devices, scientific instruments, instruments, etc., can solve the problems of missed diagnosis, obtaining test results, and complicated method operation, and achieve the effect of avoiding missed detection, ensuring specificity, and ensuring sensitivity

Inactive Publication Date: 2015-09-16
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing HCV clinical detection methods mainly include PCR detection, hepatitis C virus antibody detection and hepatitis C virus core antigen detection kit. Among the three detection methods, the PCR detection method has high sensitivity and accurate results, but the method is cumbersome to operate , the required personnel have strong technical skills, and it is not easy to promote it in various hospitals; the hepatitis C virus antibody detection reagent is the most common hepatitis C detection method in clinica

Method used

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  • Kit for combined detection of hepatitis C virus antigen and antibody through chemiluminescence
  • Kit for combined detection of hepatitis C virus antigen and antibody through chemiluminescence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] The components and concentrations of the detection reagents were

[0076] Component 1 (R1) magnetic particle reagent:

[0077] Magnetic particle and goat anti-FITC antibody conjugate 1%

[0078] Tris 12.1g / L

[0079] BSA 0.5g / L

[0080] PC-300 0.1%

[0081] Component 2 (R2) FITC labeling reagent:

[0082] FITC-labeled HCV-cAg antibody 1%

[0083] FITC-labeled NS3 protein 1%

[0084] FITC-labeled NS4 protein 1%

[0085] FITC-labeled NS5 protein 1%

[0086] Tris 12.1g / L

[0087] BSA 0.5g / L

[0088] PC-300 0.1%

[0089] Component 3 (R3) AP Labeling Reagent:

[0090] AP labeled HCV-cAg antibody 1%

[0091] AP-tagged NS3 protein 1%

[0092] AP-tagged NS4 protein 1%

[0093] AP-tagged NS5 protein 1%

[0094] Tris 12.1g / L

[0095] BSA 0.5g / L

[0096] PC-300 0.1%

[0097] Component 4 (R4) sample diluent:

[0098] Disodium hydrogen phosphate 1.974g / L

[0099] Potassium dihydrogen phosphate 0.2245g / L

[0100] BSA 0.5g / L

[0101] PC-300 0.1%

[0102] ...

Embodiment 2

[0135] The routine hepatitis C virus core antigen detection kit (enzyme-linked immunoassay) is selected from the American company Ortho, and the detection operation steps are as follows:

[0136] (1) Equilibration: Take out the kit and samples from the refrigerated environment, and place them at 15-30°C for 30 minutes;

[0137] (2) Dosing: Shake all reagents well before use, pay attention to no residual crystals in the bottle, and dilute the concentrated washing solution 20 times with distilled or deionized water;

[0138] (3) Adding samples: 2 wells each for blank, negative and positive controls are required for each test. Add 200 μl sample diluent to the blank well, first add 100 μl sample diluent to the other wells, then add 100 μl negative control to each well of the negative control well, add 100 μl positive control to each well of the positive control well, and add 100 μl sample to be tested to each well;

[0139] (4) Incubation: After mixing evenly, stick on the sealing ...

Embodiment 3

[0146] The accuracy of the kit for combined detection of hepatitis C virus antigen-antibody by chemiluminescence method was analyzed, and the kit of Ortho Company in the United States was used as the gold standard to detect 40 cases of clinical samples, and the results were compared. The detection method is as in Example 1 , embodiment 2, if there are differences in the test results, send a third-party inspection agency to verify by using the PCR detection method, and the test results are as shown in Table 1:

[0147] Table 1 Accuracy test results

[0148]

[0149] After comparative testing, the test results of No. 8 and No. 24 samples were abnormal. After the third-party test verification, the test results of Example 1 (the present invention) were more accurate. The two abnormal samples were mainly due to autoimmune antibodies, while The concentration of hepatitis C virus core antigen is relatively low, resulting in missed detection. According to the detection result, it ...

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Abstract

The invention provides a kit for combined detection of hepatitis C virus antigen and antibody through chemiluminescence. According to the invention, the principles of chemiluminescence are utilized; anti-FITC-coated magnetic particles are bonded with an FITC-coated reagent and then undergo immunoreaction with a detected object; after immunoreaction of an AP-labeled substance, an immunoreaction chain (as shown in a figure 1 which is described in the specification) is constructed; luminous sensitivity of a substrate catalyzed by AP is far higher than enzyme immunoassay development, so reaction sensitivity is enhanced; meanwhile, FITC is used to simultaneously marking antigen and antibody, so hepatitis C virus antibody and core antigen are detected at the same time. The kit provided by the invention can more accurately detect hepatitis C, is free of leak detection, achieves the effect of early discovery and has a high application value in prevention of hepatitis C.

Description

Technical field [0001] The invention is the field of hepatitis C testing technology, and it involves a hepatitis C virus antigen-antibody joint testing kit (chemical light emitting method). Background technique [0002] Hepatitis C is a global infectious disease caused by Hepatitis C Virus (HCV) infection.It is estimated that there are currently 1700 million hepatitis C virus infections in the world, and the hepatitis C infection rate in my country is about 30%, and currently there are at least 4,000 to 60 million patients with hepatitis C.Among them, 80 ~ 85%of the infected people will develop into chronic hepatitis C, of which 20%can develop into liver fibrosis, and eventually 4-5%of patients with liver cellular liver cancer occurred in patients with hepatitis, and the harm is very serious. [0003] At present, there is no special effects therapeutic drug for hepatitis C virus. The development of hepatitis C vaccine is also difficult to change due to the rapid mutation of HCV, ...

Claims

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Application Information

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IPC IPC(8): G01N33/576
CPCG01N33/5767G01N33/56983G01N2333/186
Inventor 魏海明王进谭柏清王春艳王绮
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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