采用反转录-聚合酶链反应结合限制性内切酶分析快速鉴定传染性法氏囊病病毒的“非常毒力”株。

Rapid identification of "very virulent" strains of infectious bursal disease virus by reverse transcription-polymerase chain reaction combined with restriction enzyme analysis.

机构信息

Institut fur Virologie, Veterinarmedizinische Fakultat, Universitat Leipzig, An den Tierkliniken 29, D-04103 Leipzig, Germany.

出版信息

Avian Pathol. 2001 Feb;30(1):55-62. doi: 10.1080/03079450020023203.

DOI:10.1080/03079450020023203

PMID:19184874

Abstract

A reverse transcription-polymerase chain reaction (RT-PCR) combined with restriction enzyme analysis (REA) was developed for differentiation of classical virulent (cv) and very virulent (vv) strains of infectious bursal disease virus (IBDV). The VP2 gene was used for primer annealing to amplify the hypervariable region. An amplification product was obtained with serotype 1 and serotype 2 strains of IBDV.The restriction enzymes SacI and BspMI were used to identify and to differentiate the serotype 1 strains: SacI only cleaved cvIBDV RT-PCR products, whereas products obtained with vvIBDV strains were only cleaved with BspMI; serotype 2 strain products were not cleaved by either of these restriction enzymes. RT-PCR combined with REA as described in this study is thus suitable to detect IBDV of both serotypes and to distinguish rapidly between cvIBDV,vvIBDV and serotype 2 IBDV.Our investigation of a total of 11 different IBDV isolates and available nucleotide sequence data of other isolates indicate that the application of this protocol might be a useful tool for the rapid identification of vvIBDV isolates, a prerequisite for the effective control of this economically important virus infection of commercial poultry.

摘要

建立了反转录-聚合酶链反应（RT-PCR）结合限制性内切酶分析（REA）方法，用于区分传染性法氏囊病病毒（IBDV）的经典强毒（cv）和超强毒（vv）株。使用 VP2 基因进行引物退火以扩增高变区。使用 IBDV 的血清型 1 和血清型 2 菌株获得了扩增产物。SacI 和 BspMI 两种限制酶用于鉴定和区分血清型 1 株：SacI 仅切割 cvIBDV RT-PCR 产物，而 vvIBDV 株的产物仅被 BspMI 切割；血清型 2 株的产物不受这两种限制酶中的任何一种切割。因此，本研究中描述的 RT-PCR 结合 REA 适合检测两种血清型的 IBDV，并能快速区分 cvIBDV、vvIBDV 和血清型 2 IBDV。我们对总共 11 种不同的 IBDV 分离株进行了研究，并对其他分离株的可用核苷酸序列数据进行了分析，表明该方案的应用可能是快速鉴定 vvIBDV 分离株的有用工具，这是有效控制这种经济上重要的商业家禽病毒感染的前提。