THE DETECTION OF TOMATO SPOTTED WILT VIRUS USING THE POLYMERASE CHAIN-REACTION

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.