Mumford R A, Barker I, Wood K R
School of Biological Sciences, University of Birmingham, Edgbaston, UK.
J Virol Methods. 1994 Mar;46(3):303-11. doi: 10.1016/0166-0934(94)90002-7.
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.
基于聚合酶链反应（PCR）的检测方法用于从感染的植物材料中提取的总RNA中检测多个英国番茄斑萎病毒（TSWV）分离株。提取物进行逆转录，然后使用针对L RNA片段276个碱基对片段的寡核苷酸引物通过PCR扩增所得的cDNA。检测产物在琼脂糖凝胶上进行电泳，并用溴化乙锭染色进行可视化。通过测序确认了所产生产物的病毒来源，获得的数据与先前发表的L RNA序列数据具有非常高的同源性。讨论了与现有检测方法相比，RT-PCR检测方法的特异性和灵敏度。
