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基于聚合酶链反应(PCR)对来自加拿大萨斯喀彻温省胎儿三毛滴虫分离株保守和可变DNA序列的研究。

PCR-based study of conserved and variable DNA sequences of Tritrichomonas foetus isolates from Saskatchewan, Canada.

作者信息

Riley D E, Wagner B, Polley L, Krieger J N

机构信息

Department of Urology, University of Washington School of Medicine, Seattle 98195, USA.

出版信息

J Clin Microbiol. 1995 May;33(5):1308-13. doi: 10.1128/jcm.33.5.1308-1313.1995.

Abstract

The protozoan parasite Tritrichomonas foetus causes infertility and spontaneous abortion in cattle. In Saskatchewan, Canada, the culture prevalence of trichomonads was 65 of 1,048 (6%) among 1,048 bulls tested within a 1-year period ending in April 1994. Saskatchewan was previously thought to be free of the parasite. To confirm the culture results, possible T. foetus DNA presence was determined by the PCR. All of the 16 culture-positive isolates tested were PCR positive by a single-band test, but one PCR product was weak. DNA fingerprinting by both T17 PCR and randomly amplified polymorphic DNA PCR revealed genetic variation or polymorphism among the T. foetus isolates. T17 PCR also revealed conserved loci that distinguished these T. foetus isolates from Trichomonas vaginalis, from a variety of other protozoa, and from prokaryotes. TCO-1 PCR, a PCR test designed to sample DNA sequence homologous to the 5' flank of a highly conserved cell division control gene, detected genetic polymorphism at low stringency and a conserved, single locus at higher stringency. These findings suggested that T. foetus isolates exhibit both conserved genetic loci and polymorphic loci detectable by independent PCR methods. Both conserved and polymorphic genetic loci may prove useful for improved clinical diagnosis of T. foetus. The polymorphic loci detected by PCR suggested either a long history of infection or multiple lines of T. foetus infection in Saskatchewan. Polymorphic loci detected by PCR may provide data for epidemiologic studies of T. foetus.

摘要

原生动物寄生虫胎儿三毛滴虫可导致牛的不育和自然流产。在加拿大萨斯喀彻温省,截至1994年4月的1年时间内,在检测的1048头公牛中,滴虫的培养阳性率为1048头中的65头(6%)。萨斯喀彻温省以前被认为没有这种寄生虫。为了证实培养结果,通过聚合酶链反应(PCR)确定是否存在胎儿三毛滴虫DNA。所有检测的16株培养阳性分离株通过单带试验PCR呈阳性,但有一个PCR产物较弱。通过T17 PCR和随机扩增多态性DNA PCR进行的DNA指纹分析揭示了胎儿三毛滴虫分离株之间的遗传变异或多态性。T17 PCR还揭示了保守位点,这些位点将这些胎儿三毛滴虫分离株与阴道毛滴虫、多种其他原生动物以及原核生物区分开来。TCO - 1 PCR是一种旨在对与高度保守的细胞分裂控制基因5'侧翼同源的DNA序列进行采样的PCR检测方法,在低严谨度下检测到遗传多态性,在高严谨度下检测到一个保守的单一位点。这些发现表明,胎儿三毛滴虫分离株表现出可通过独立PCR方法检测到的保守遗传位点和多态性位点。保守和多态性遗传位点可能对改进胎儿三毛滴虫的临床诊断有用。通过PCR检测到的多态性位点表明萨斯喀彻温省存在长期感染史或多条胎儿三毛滴虫感染途径。通过PCR检测到的多态性位点可能为胎儿三毛滴虫的流行病学研究提供数据。

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