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
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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.
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(4):1578-1589.
Xue, Xianghong; Zheng, Xuexing; Liang, Hongru; Feng, Na; Zhao, Yongkun; Gao, Yuwei; Wang, Hualei; Yang, Songtao; Xia, Xianzhu. 2014. "Generation of Recombinant Rabies Virus CVS-11 Expressing eGFP Applied to the Rapid Virus Neutralization Test." Viruses 6, no. 4: 1578-1589.