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

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

1,2,†
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1,3,†
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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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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.
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 3.0).

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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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