The detection of tomato spotted wilt virus using the polymerase chain reaction.
A polymerase chain reaction (PCR) based assay was used to detected a number of UK tomato spotted wilt virus (TSWV) isolates in total RNA extractions made from infected plant material. Extracts were reverse transcribed and the resultant cDNA amplified by PCR, using oligonucleotide primers specific for a 276 base pair fragment of the L RNA segment. Assay products were electrophoresed on agarose gels and visualised by ethidium bromide staining. The viral origin of the product produced was confirmed by sequencing, with the data obtained having very high homology with previously published L RNA sequence data. The specificity and sensitivity of the RT-PCR assay, in comparison with existing tests, is discussed.
Mumford
R A
RA
School of Biological Sciences, University of Birmingham, Edgbaston, UK.
Barker
I
I
Wood
K R
KR
D001483:Base Sequence; D002044:Bunyaviridae Infections / Q000175:diagnosis*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D017865:Tospovirus / Q000235:genetics / Q000302:isolation & purification*
D012367:RNA, Viral