B. Liess et al., AN IMMUNOPLAQUE ASSAY DISTINGUISHING BETWEEN CYTOPATHOGENIC AND NONCYTOPATHOGENIC BIOTYPES OF BOVINE VIRAL DIARRHEA VIRUS, Journal of veterinary medicine. Series B, 40(2), 1993, pp. 89-96
Cytopathogenic (cp) and noncytopathogenic (ncp) biotypes of bovine vir
us diarrhoea virus (BVDV) could be clearly distinguished by hollow pla
ques with peripheral immunostaining in infected fetal calf kidney (FCK
) monolayers cultures or the appearance of homogenously stained immuno
plaques (IPs), respectively. Of 70 BVDV isolates formerly classified a
s cytopathogenic according to microscopic observations in FCK cell cul
tures, 46 produced IPs of both types while the remaining 24 isolates i
nduced only the IP type characteristic of the ncp BVDV biotype. The sa
me was true for 24 additional isolates classified by light microscopy
as ncp after ten blind passages in FCK cell cultures. Thus it appeared
that in 1/3 of the cases changes in FCK cell cultures had been former
ly misinterpreted as BVDV induced cytopathic effects (CPE). Of the 46
isolates which produced both ncp and cp immunoplaques 5 BVDV isolates
were selected and successfully subjected to cloning. Clones of the cp
biotype were developed by picking native plaques from unfixed FCK cell
monolayers. In case of ncp IP, cloning was attempted by picking rando
mly in the spaces between visible native plaques. Pairs of clones from
each of the field isolates proved to consist of pure plaque populatio
ns of cp BVDV and ncp BVDV biotypes, respectively, at the dilution of
viral suspension which allowed the classification of a single IP accor
ding to morphology. Analysis of the clones using 15 monoclonal antibod
ies against the major viral glycoprotein revealed far-reaching antigen
ic homogeneity for each pair with only few but distinct exceptions.