Xu L, Hampton R O
Department of Botany and Plant Pathology, Oregon State University, Corvallis, USA.
Arch Virol. 1996;141(10):1961-77. doi: 10.1007/BF01718207.
Partial nucleotide sequences of selected isolates of bean common mosaic virus (BCMV) and bean common mosaic necrosis virus (BCMNV) were determined. Based on these sequences and previously published sequence data, a reverse transcription, polymerase chain reaction (RT-PCR) in combination with restriction endonuclease analyses, was developed for molecular detection of BCMV, BCMNV and some viral pathogroups (PG). Specific detection of the two viruses was accomplished by constructing two virus-specific primer pairs that amplified a PCR product specific for each virus. By application of RT-PCR, four BCMV-PG-V isolates were differentiated from isolates of BCMV pathogroups I, II, IV and VII. Distinction of two BCMNV pathogroups (PG-III and PG-VI) was achieved by restriction enzyme XbaI digestion of BCMNV PCR products. However, no combination of tested restriction enzymes distinguished all five recognized BCMV pathogroups. A primer pair Dts/Uny15 proved to be specific for BCMV pathogroup PG-V. Thus, by a combination of RT-PCR and restriction enzyme analyses, it was possible to differentiate both viruses, two pathogroups of BCMNV, and one pathogroup of BCMV from the others.
测定了菜豆普通花叶病毒(BCMV)和菜豆普通花叶坏死病毒(BCMNV)部分分离物的核苷酸序列。基于这些序列和先前发表的序列数据,开发了一种逆转录聚合酶链反应(RT-PCR)并结合限制性内切酶分析,用于BCMV、BCMNV和一些病毒致病型(PG)的分子检测。通过构建两对病毒特异性引物对来实现对这两种病毒的特异性检测,这两对引物对可扩增出每种病毒特有的PCR产物。通过应用RT-PCR,将四个BCMV-PG-V分离物与BCMV致病型I、II、IV和VII的分离物区分开来。通过对BCMNV PCR产物进行限制性内切酶XbaI消化,实现了对两个BCMNV致病型(PG-III和PG-VI)的区分。然而,所测试的限制性酶的任何组合都无法区分所有五个公认的BCMV致病型。引物对Dts/Uny15被证明对BCMV致病型PG-V具有特异性。因此,通过RT-PCR和限制性酶分析相结合,可以将这两种病毒、两个BCMNV致病型和一个BCMV致病型与其他类型区分开来。