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.
Highlights
Rabies is a fatal zoonotic infectious disease caused by the rabies virus (RABV) that affects numerous warm-blooded mammals worldwide [1]
To verify the rescued RABV rCVS-11-enhanced green fluorescent protein (eGFP) strain replication and expression of eGFP as an RABV structural protein, Baby hamster kidney (BHK)-21 cells infected with rCVS-11-eGFP were immunostained with
To confirm that the rCVS-11-eGFP strain was derived from the full-length plasmid pCVS-11-eGFP, BHK-21 cells infected with the rCVS-11-eGFP strain were used to perform
Summary
Rabies is a fatal zoonotic infectious disease caused by the rabies virus (RABV) that affects numerous warm-blooded mammals worldwide [1]. Dogs and cats are the major reservoir of human rabies in most parts of the developing world [5], and the vast majority of human rabies victims become infected by dog bites [6,7]. 70% immunization of the dog population can efficiently block RABV transmission [4,8], and that virus-neutralizing antibody plays an important role in protecting against RABV [9,10]. Quantification of the VNA levels against RABV is essential for the timely monitoring of immunization coverage in dog and cat populations and to evaluate the immunity effectiveness of RABV vaccines
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