Blanco-Urgoiti B, Sánchez F, Dopazo J, Ponz F
Laboratorio de Biología Molecular y Virología Vegetal, CIT-INIA, Madrid, Spain.
Arch Virol. 1996;141(12):2425-42. doi: 10.1007/BF01718641.
Potato virus Y (PVY) isolates have been classified into genetic strains by a host-independent criterion using a molecular typing method. The method used extracts from infected tissue, and included immunocapture-RT-PCR-RFLP analysis using 5 different restriction endonucleases (Dde I, Eco RV, Hinf I, Rsa I and Taq I). Genetic distances between the different PVY "restrictotypes" were calculated and used to define the PVY genetic strains. Three main clusters were found: PVYO, PVYN, and non-potato PVY (PVYNP), in good agreement with classical PVY strain definitions that combine different biological criteria. Our approach was incomparably quicker and more reliable and reproducible than biotyping. The potential of this approach for very quick, simple and automatable molecular epidemiological studies is discussed.
马铃薯Y病毒(PVY)分离株已通过分子分型方法,采用与宿主无关的标准被分类为不同的遗传株系。所使用的方法是从受感染组织中提取样本,并包括使用5种不同的限制性内切酶(Dde I、Eco RV、Hinf I、Rsa I和Taq I)进行免疫捕获RT-PCR-RFLP分析。计算了不同PVY“限制型”之间的遗传距离,并用于定义PVY遗传株系。发现了三个主要聚类:PVYO、PVYN和非马铃薯PVY(PVYNP),这与结合不同生物学标准的经典PVY株系定义高度一致。我们的方法比生物分型无比快速、可靠且可重复。讨论了这种方法在非常快速、简单且可自动化的分子流行病学研究中的潜力。