Fast test strip of vesicular stomatitis virus (VSV) and preparation method thereof

A technology of vesicular stomatitis and detection test paper, applied in the biological field, can solve the problems of unavailable rapid diagnosis, non-specific reaction, and complicated test of diagnostic methods, and achieve the advantages of less reagents and samples, strong specificity, and high sensitivity Effect

Inactive Publication Date: 2014-03-26
花群义
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Problems solved by technology

Among them, the two methods of virus neutralization test (VN) and complement fixation test (CF) are complex, cumbersome, time-consuming, unsafe, need to be carried out in a biosafety level 3 (P3) laboratory, and sometimes produce non-specificity. Reaction, the application is limited, and other diagnostic methods also have their own limitations and cannot be used for rapid diagnosis

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  • Fast test strip of vesicular stomatitis virus (VSV) and preparation method thereof

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Embodiment Construction

[0022] The following examples are further explanations and illustrations of the present invention, and do not constitute any limitation to the present invention.

[0023] 1. Preparation process of monoclonal antibody against vesicular stomatitis virus:

[0024] (1) BHK-21 cells were grown into a monolayer and inoculated with vesicular stomatitis virus (VSV), and reacted at a temperature of 37° C. for 1 hour. Serum-free basal medium was added to continue culturing, and the virus was harvested when the cytopathic pathology (CPE) reached more than 75% after 2-3 days. The harvested virus liquid was repeatedly frozen and thawed 3 times, centrifuged at 8000 rpm for 30 minutes, and the supernatant was taken. Ultracentrifuge at 28,000 rpm at 4°C for 3 hours. Dissolve the precipitate with 1 / 100 of the original volume of PBS, then use 20%, 60% (w / v) discontinuous sucrose density gradient at 20,000 rpm, ultracentrifuge at 4°C for 3 hours, and harvest the purified virus band as the immu...

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Abstract

The invention relates to a fast test strip of vesicular stomatitis virus (VSV) and a preparation method thereof. The test strip comprises a base plate (1), a sample absorption pad (2) of a monoclonal antibody absorbed with VSV labelled with colloidal gold, a nitrocellulose membrane (3) provided with a detection line (4) and a control line (5) and an absorbent pad (6), and the absorption pad (2), the nitrocellulose membrane (3) and the absorbent pad (6) are sequentially arranged in the base plate (1) along with the length direction. The method comprises the steps of: a, preparing the VSV monoclonal antibody; b, burning colloidal gold and preparing a monoclonal antibody to be labelled; c, labelling the antibody with colloidal gold and purifying the antibody; d, absorbing the purified antibody labelled with colloidal gold on a multiporous fiber sample absorption pad; e, forming the detection line and the control line coated or sprayed by VSV and protein A; and f, arranging the sample absorption pad, the nitrocellulose membrane and the absorbent pad on the plate of the base plate sequentially and forming the test strip. Without special equipment and special technicians, the VSV can be directly and fast detected.

Description

【Technical field】 [0001] The invention relates to the field of biotechnology, in particular to a rapid detection test strip for detecting vesicular stomatitis virus antigen and a manufacturing method thereof. 【Background technique】 [0002] Vesicular stomatitis is a major zoonotic disease caused by vesicular stomatitis virus (VSV) belonging to the family Rhabdoviridae and genus Vesiculovirus. It includes Indiana (Indiana, IN type) and New Jersey (New Jersey, NJ type) two serotypes. Cattle, pigs, horses, sheep, and many other wild animals can be infected, and humans can develop flu-like and dengue-like symptoms after infection. The clinical symptoms are not easy to distinguish from foot-and-mouth disease (FMD), and the existence and prevalence of the disease directly affect the health of humans and animals. The genome code of vesicular stomatitis virus contains five kinds of viral proteins, including N, NS, M, G, and L. Among them, the N protein is the main protein of the n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/577G01N33/532
Inventor 花群义阮周曦杨俊兴杨云庆曾少灵张彩虹孙洁吕建强秦智锋陶虹
Owner 花群义
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