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用于鉴定中国大陆旋毛虫分离株的核糖体DNA的聚合酶链反应-单链构象多态性分析

PCR-SSCP of rDNA for the identification of Trichinella isolates from mainland china.

作者信息

Gasser R B, Zhu X Q, Monti J R, Dou L, Cai X, Pozio E

机构信息

Department of Veterinary Science, The University of Melbourne, Victoria, Australia.

出版信息

Mol Cell Probes. 1998 Feb;12(1):27-34. doi: 10.1006/mcpr.1997.0142.

Abstract

A polymerase chain reaction (PCR)-based single strand conformation polymorphism (SSCP) analysis of the expansion segment 5 (domain IV) of the large subunit of ribosomal DNA was employed to characterize seven isolates of Trichinella from China (A-G), including six of pig origin from regions in Dengxian (A), Tianjin (B), Harbin (D), Baoshan (E), Xinye (F) and Xian (G), and one of canine origin from Changchun (C). Isolates A, D, E, F and G were classified as Trichinella spiralis based on SSCP patterns, while the patterns for isolates B and C were consistent with those of Trichinella nativa or Trichinella T6. The results were supported by random amplified polymorphic DNA (RAPD) analysis using five decamer primers and were in accordance with ecological information for the isolates. Single strand conformation polymorphism results also allowed the direct display of mutational sequence variation in the expansion segment among the five isolates of T. spiralis from China, indicating its usefulness for studying population variation within that species.

摘要

采用基于聚合酶链反应(PCR)的核糖体DNA大亚基扩展片段5(结构域IV)单链构象多态性(SSCP)分析,对来自中国的7株旋毛虫分离株(A - G)进行特征分析,其中包括6株猪源分离株,分别来自邓县(A)、天津(B)、哈尔滨(D)、宝山(E)、新野(F)和西安(G)地区,以及1株长春犬源分离株(C)。根据SSCP图谱,分离株A、D、E、F和G被归类为旋毛形线虫,而分离株B和C的图谱与本地旋毛虫或旋毛虫T6一致。使用5种十聚体引物进行的随机扩增多态性DNA(RAPD)分析支持了该结果,且与分离株的生态信息相符。单链构象多态性结果还能直接显示中国5株旋毛形线虫分离株扩展片段中的突变序列变异,表明其在研究该物种内群体变异方面的有用性。

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