Ra. Mumford et al., THE DETECTION OF TOMATO SPOTTED WILT VIRUS USING THE POLYMERASE CHAIN-REACTION, Journal of virological methods, 46(3), 1994, pp. 303-311
A polymerase chain reaction (PCR) based assay was used to detected a n
umber of UK tomato spotted wilt virus(TSWV) isolates in total RNA extr
actions made from infected plant material. Extracts were reverse trans
cibed and the resultant cDNA amplified by PCR, using oligonucleotide p
rimers specific for a 276 base pair fragment of the L RNA segment. Ass
ay products were electrophoresed on agarose gels and visualised by eth
idium bromide staining. The viral origin of the product produced was c
onfirmed by sequencing, with the data obtained having very high homolo
gy with previously published L RNA sequence data. The specificity and
sensitivity of the RT-PCR assay, in comparison with existing tests, is
discussed.