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Enzyme linked immunoreaction reagent kit for detecting rabies virus

An enzyme-linked immunosorbent reagent, rabies virus technology, applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of inability to meet large-scale rapid diagnosis, increase the risk of operators, pollution, etc., and achieve suitable for large-scale investigation The effect of shortening the work and incubation time, and the operation is simple and fast

Inactive Publication Date: 2007-03-14
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Moreover, the above methods require professional experimental techniques and equipment, and often take several days or even weeks to obtain results, which cannot meet the needs of large-scale rapid diagnosis at all.
Although there are also research reports on rabies ELISA detection kits in China, the antigens used in their research all use inactivated rabies virus. Since rabies virus is easily transmitted through damaged skin and mucous membranes, it undoubtedly increases the risk of operators.
Moreover, if the inactivation of rabies virus is not complete during the production process, it will cause a wider range of pollution and spread

Method used

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  • Enzyme linked immunoreaction reagent kit for detecting rabies virus
  • Enzyme linked immunoreaction reagent kit for detecting rabies virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1, the preparation of the ELISA kit that detects rabies virus

[0027] 1. Preparation of enzyme-linked immunosorbent assay kit for detection of rabies virus

[0028] The ELISA kit of rabies virus consists of:

[0029] 1) A microtiter plate coated with rabies virus N protein

[0030] 2) Sample diluent: 10ml of a solution containing 2.1% NaCl and 4% PEG6000

[0031] 3) Positive serum 0.5ml

[0032] 4) Negative serum 0.5ml

[0033] 5) Horseradish peroxidase (HRP) labeled anti-dog IgG 10ml

[0034] 6) Concentrated washing liquid: 20ml

[0035] 7) Chromogenic agent: Chromogenic substrate A: 5ml of 3.3'-5.5'-tetramethylbenzidine solution; Chromogenic substrate B: 5ml of hydrogen peroxide solution;

[0036] 9) Stop solution: 5ml of 2mol / L sulfuric acid solution diluted with distilled water

[0037] The preparation method of above-mentioned each component is as follows:

[0038] 1. Preparation of an ELISA plate coated with rabies virus N protein

[0039] 1) ...

Embodiment 2

[0080] Embodiment 2, the effect experiment of the ELISA kit that detects rabies virus

[0081] 1. Kit specific detection:

[0082] Collect 20 rabies virus-negative sera and 20 rabies virus-positive sera respectively, and use the fluorescent antibody virus neutralization test (FAVN) to determine, the specific operation refers to the literature report (Cliquet F, Aubert M, Sagné L. (FAVN) for the quantitation of rabies-neutralising antibody.Journal of ImmunologicalMethods, 1998, 212:79-87) the enzyme-linked immunoassay kit detection of the detection rabies virus prepared by the method of Example 1, the result shows that there is Eighteen were positive, and 20 of the negative sera were negative, indicating a 95% detection accuracy.

[0083] 2. Sensitivity detection: the standard rabies virus positive serum (Changchun Veterinary Research Institute of Military Medical Sciences) is gradiently diluted with the sample diluent prepared in Example 1, and the ELISA kit for detecting rab...

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Abstract

The disclosed enzyme linked immunity reagent for rabies virus comprises: the enzyme mark plate covered by rabies virus N protein, and the enzyme label as enzyme-labeled antibody. This invention has specificity up to 95%, sensitivity up to 0.25IU / ml, and precision (coefficient of variation C .V) up to 8.68%, and can complete the detection in one hour.

Description

technical field [0001] The invention relates to an ELISA kit for detecting rabies virus. Background technique [0002] Rabies is a zoonotic infectious disease of central nervous system infection caused by rabies virus, and the mortality rate after its onset is 100%. The death toll of rabies in my country ranks second in the world, second only to India. The case fatality rate and death toll of rabies rank first among Class A and Class B infectious diseases in my country. [0003] According to the "National Rabies Surveillance Program", the domestic detection of rabies mainly adopts ① pathogen detection method: including immunofluorescence method for antigen detection and rapid rabies enzyme-linked immunosorbent assay for antigen detection; ② RT-PCR nucleic acid detection method; ③ virus isolation method: including Isolation of virus by cell culture method and inoculation of suckling mice; and ④ Antibody detection method: including specific antibody detection and neutralizin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/547G01N33/53G01N21/78C12Q1/68
Inventor 刘文军杨利敏
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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