Eh. Venter et al., DETECTION OF BLUETONGUE VIRUS AND AFRICAN HORSESICKNESS VIRUS IN CO-INFECTED CELL-CULTURES WITH NS1 GENE PROBES, Onderstepoort journal of veterinary research, 62(4), 1995, pp. 217-222
The serogroup specificity of the bluetongue virus (BTV) NS1 and VP3 ge
ne probes was confirmed by means of northern blot hybridization. Under
high-stringency conditions both probes hybridized to 22 BTV serotypes
(18 South African serotypes, BTV3 from Cyprus and BTV16 from Pakistan
) but not to serotypes that originate from Australia and India. Furthe
rmore, NS1 gene probes of BTV and African horsesickness virus (AHSV) w
ere used in a dot-spot in site hybridization procedure to differentiat
e between BTV and AHSV in co-infected cell cultures. The method detect
s viral RNA directly in glutaraldehyde-fixed infected cell cultures wi
thout prior nucleic-acid extraction or purification. AHSV could be det
ected in cells infected with AHSV at a multiplicity of infection of 10
(-4) PFU/cell in the presence of a hundredfold excess of co-infecting
BTV. The method may have an application in epidemiological surveys to
detect different orbiviruses in the same Culicoides population.