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[利用聚合酶链反应改进鼠疫微生物菌株检测方法]

[Improvement of a method for detecting of strains of the plague microbe using polymerase chain reaction].

作者信息

Kulichenko A N, Norkina O V, Gintsburg A L, Popov Iu A, Ddrozdov I G

出版信息

Genetika. 1994 Feb;30(2):167-71.

PMID:8045376
Abstract

Three pairs of oligonucleotide primers, complementary to nucleotide sequences of Yersinia pestis plasmids (pPst, 9.5 kb; pCad, 70 kb; pFra, 95 kb), were used in polymerase chain reaction for high-sensitive specific detection of plague pathogen. Primer pairs P1,P2,C1,C2, and F1,F2 were used to amplify fragments of pla gene (plasmid pPst), yop1 gene (plasmid pCad of plague microbe), and caf1 gene (plasmid pFra), respectively. The method developed enables specific detection of strains from all natural loci in Russia and contiguous states as well as strains of oceanic origin. Sensitivity of method is 50-100 CFU/ml. Primer sequences enable to amplify gene fragments, located in three own plasmides of plague microbe, in the same reaction mixture. The method offers identification of plague microbe and determination of its virulence and epidemic significance.

摘要

三对与鼠疫耶尔森氏菌质粒(pPst,9.5 kb;pCad,70 kb;pFra,95 kb)核苷酸序列互补的寡核苷酸引物,用于聚合酶链反应,以高灵敏度特异性检测鼠疫病原体。引物对P1、P2、C1、C2和F1、F2分别用于扩增pla基因(质粒pPst)、yop1基因(鼠疫微生物的质粒pCad)和caf1基因(质粒pFra)的片段。所开发的方法能够特异性检测来自俄罗斯所有天然疫源地以及毗邻国家的菌株以及海洋来源的菌株。该方法的灵敏度为50 - 100 CFU/ml。引物序列能够在同一反应混合物中扩增位于鼠疫微生物三个自身质粒中的基因片段。该方法可用于鼠疫微生物的鉴定及其毒力和流行意义的确定。

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