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Novel neutralizing antibody detection method based on rescued recombinant attenuated rabies virus

An antibody detection and rabies technology, applied in recombinant DNA technology, inactivation/attenuation, introduction of foreign genetic material using vectors, etc. Clear point, simple operation steps, weak virulence effect

Inactive Publication Date: 2014-12-10
GUANGXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, RFFIT and FAVN use the rabies virus CVS-11 strain as the standard challenge strain, which has strong virulence and potential safety hazards; at the same time, the operation is more complicated, requiring strict laboratory conditions and skilled operators; And expensive specific anti-rabies virus primary antibody and FITC-labeled secondary antibody are not conducive to the detection of rabies virus neutralizing antibody level at the grassroots level

Method used

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  • Novel neutralizing antibody detection method based on rescued recombinant attenuated rabies virus
  • Novel neutralizing antibody detection method based on rescued recombinant attenuated rabies virus
  • Novel neutralizing antibody detection method based on rescued recombinant attenuated rabies virus

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Embodiment 1

[0052] 1. Preparation of materials

[0053] Rabies virus strains RC-HL, pRC-HL(+), helper plasmids p-N, p-P and p-L were donated by Gifu University, Japan, and BSR-T7 cells, BHK-21 cells, NA cells and peGFP-N1 vectors were preserved by the applicant. Rabies virus P polyclonal antibody was prepared and preserved by the applicant, rabies vaccine (Merial, France) immunized and non-immunized canine serum samples were provided by Guangxi Animal Disease Prevention and Control Center, and the serum was from Longan County Animal Disease Prevention and Control Center. 4-week-old Kunming mice were purchased from the Experimental Animal Center of Guangxi University of Traditional Chinese Medicine.

[0054] 2. Design and synthesis of primers for constructing recombinant virus

[0055] According to the P and M sequences and eGFP sequences of RC-HL strains in GenBank, three pairs of overlapping PCR primers were designed by using Oligo6.0 software. Primers P-F and P-R amplify the P-1 gene,...

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Abstract

The invention discloses a novel neutralizing antibody detection method based on a rescued recombinant attenuated rabies virus. The inventor rescues a recombinant virus rRV-eGFP with green fluorescent protein by utilization of a reverse genetics technology and improves the internationally accepted RFFIT method for valence detection of a neutralizing antibody against rabies virus by adoption of the recombinant virus, thus developing an effective modified RFFIT-eGFP method. The novel neutralizing antibody detection method improves detection effects of the RFFIT method, and has characteristics of good consistency, high specificity, high safety, no need of expensive primary antibodies or FITC-labeled second antibodies, capability of being more rapid, simpler and more convenient, and the like when compared with the standard RFFIT method. The novel neutralizing antibody detection method overcomes the problems, namely use of a challenge virus standard strain that is a CVS-11 strain with high virulence, long consumed time, and the like of the traditional RFFIT method.

Description

technical field [0001] The invention belongs to the technical field of detection of rabies virus neutralizing antibody, and in particular relates to a novel neutralizing antibody detection method based on rescuing recombinant rabies virus attenuated virus. Background technique [0002] Rabies is a worldwide zoonotic disease caused by neurotropic rabies virus (RV). It is an ancient disease that can infect all warm-blooded animals. About 55,000 people die from rabies every year in the world (WHO data), mainly concentrated in some backward countries and regions such as Asia and Africa. The number of rabies patients in my country is showing an increasing trend year by year, second only to India, and rabies has become one of the public health issues of concern in our country. Once the symptoms of rabies appear, the fatality rate is close to 100%, and there is currently no effective treatment. Dogs are the main source of human rabies infection in my country, accounting for 85% t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C12N7/04C12N15/62C12N15/63
Inventor 罗廷荣唐海波钟一治韦显凯陆专灵李晓宁廖素环梁晶晶
Owner GUANGXI UNIV
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