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Viruses 2014, 6(4), 1578-1589; doi:10.3390/v6041578

Generation of Recombinant Rabies Virus CVS-11 Expressing eGFP Applied to the Rapid Virus Neutralization Test

1,2,*  and 1,2,3,*
1 Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China 2 College of Veterinary Medicine, Jilin University, Changchun 130062, China 3 Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China These authors contributed equally to this work.
* Authors to whom correspondence should be addressed.
Received: 6 January 2014 / Revised: 26 February 2014 / Accepted: 13 March 2014 / Published: 4 April 2014
(This article belongs to the Section Animal Viruses)
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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.
Keywords: rabies virus; eGFP; virus-neutralizing antibody (VNA); FAVN rabies virus; eGFP; virus-neutralizing antibody (VNA); FAVN
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Xue, X.; Zheng, X.; Liang, H.; Feng, N.; Zhao, Y.; Gao, Y.; Wang, H.; Yang, S.; Xia, X. Generation of Recombinant Rabies Virus CVS-11 Expressing eGFP Applied to the Rapid Virus Neutralization Test. Viruses 2014, 6, 1578-1589.

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