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Viruses. 2014 Apr 4;6(4):1578-89. doi: 10.3390/v6041578.

Generation of recombinant rabies Virus CVS-11 expressing eGFP applied to the rapid virus neutralization test.

Author information

1
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. red99.smile@163.com.
2
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. zhengxx2513@126.com.
3
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. hrliang13@126.com.
4
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. fengna0308@126.com.
5
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. zhaoyongkun1976@126.com.
6
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. gaoyuwei@gmail.com.
7
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. wangh25@hotmail.com.
8
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. yst610223@hotmail.com.
9
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. xiaxzh@cae.cn.

Abstract

The determination of levels of rabies virus-neutralizing antibody (VNA) provides the foundation for the quantitative evaluation of immunity effects. The traditional fluorescent antibody virus neutralization test (FAVN) using a challenge virus standard (CVS)-11 strain as a detection antigen and staining infected cells with a fluorescein isothiocyanate (FITC)-labeled monoclonal antibody, is expensive and high-quality reagents are often difficult to obtain in developing countries. Indeed, it is essential to establish a rapid, economical, and specific rabies virus neutralization test (VNT). Here, we describe a recombinant virus rCVS-11-eGFP strain that stably expresses enhanced green fluorescent protein (eGFP) based on a reverse genetic system of the CVS-11 strain. Compared to the rCVS-11 strain, the rCVS-11-eGFP strain showed a similar growth property with passaging stability in vitro and pathogenicity in vivo. The rCVS-11-eGFP strain was utilized as a detection antigen to determine the levels of rabies VNAs in 23 human and 29 canine sera; this technique was termed the FAVN-eGFP method. The good reproducibility of FAVN-eGFP was tested with partial serum samples. Neutralization titers obtained from FAVN and FAVN-eGFP were not significantly different. The FAVN-eGFP method allows rapid economical, specific, and high-throughput assessment for the titration of rabies VNAs.

PMID:
24714411
PMCID:
PMC4014711
DOI:
10.3390/v6041578
[Indexed for MEDLINE]
Free PMC Article
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