S. Zientara et al., DIAGNOSIS OF THE AFRICAN HORSE SICKNESS VIRUS SEROTYPE-4 BY A ONE-TUBE, ONE MANIPULATION RT-PCR REACTION FROM INFECTED ORGANS, Journal of virological methods, 46(2), 1994, pp. 179-188
A single tube reverse transcription - polymerase chain reaction (RT-PC
R) method for detection of African horse sickness virus (AHSV) in sple
nic tissues from infected horses is described. Double stranded RNA was
extracted from infected organs of hors es and used to produce complem
entary DNA (cDNA) with the two primers selected for the PCR. The 1179
bp amplified product (the segment 7 which encodes for VP 7), detected
by electrophoresis on agarose gel and ethidium bromide staining, was h
ydrolysed with eight restriction endonucleases for characterization of
the AHSV. The sensitivity of this method is discussed. Application of
the RT-PCR method should improve detection and shorten the time requi
red to confirm a clinical diagnosis of AHSV infection.