Generation of recombinant rabies Virus CVS-11 expressing eGFP applied to the rapid virus neutralization test.
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.
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. red99.smile@163.com.
Xianghong
X
Xue
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. zhengxx2513@126.com.
Xuexing
X
Zheng
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. hrliang13@126.com.
Hongru
H
Liang
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. fengna0308@126.com.
Na
N
Feng
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. zhaoyongkun1976@126.com.
Yongkun
Y
Zhao
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. gaoyuwei@gmail.com.
Yuwei
Y
Gao
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. wangh25@hotmail.com.
Hualei
H
Wang
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. yst610223@hotmail.com.
Songtao
S
Yang
Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun 130122, China. xiaxzh@cae.cn.
Xianzhu
X
Xia
D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood*; D004285:Dogs; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D006801:Humans; D009500:Neutralization Tests / Q000379:methods*; D030781:Organisms, Genetically Modified / Q000235:genetics / Q000276:immunology; D011820:Rabies virus / Q000235:genetics / Q000276:immunology*; D015203:Reproducibility of Results; D059386:Reverse Genetics; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins