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通过一组聚合酶链反应引物检测多种真核微生物中的可变DNA重复序列。

Detection of variable DNA repeats in diverse eukaryotic microorganisms by a single set of polymerase chain reaction primers.

作者信息

Riley D E, Samadpour M, Krieger J N

机构信息

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

出版信息

J Clin Microbiol. 1991 Dec;29(12):2746-51. doi: 10.1128/jcm.29.12.2746-2751.1991.

Abstract

We cloned and sequenced a variable DNA repeat from Trichomonas vaginalis, a flagellated protozoan parasite. Targeting of this repeat in the polymerase chain reaction resulted in complex and intense product patterns for a wide variety of eukaryotic microorganisms, including the pathogenic protozoan parasites T. vaginalis, Giardia lamblia, Leishmania donovani, three species of Trypanosoma, and four species of Acanthamoeba; the nonpathogenic protozoans, Paramecium tetraurelia and Tetrahymena thermophilia; and a yeast, Saccharomyces cerevisiae. Each microorganism exhibited a distinctive pattern of repeats. For example, a characteristic pattern was exhibited by six clinical T. vaginalis isolates. Eight G. lamblia isolates exhibited either one of two characteristic pattern types. There was no reaction with human DNA or DNA from the prokaryotes Ureaplasma urealyticum and Mycoplasma hominis. This approach may facilitate detection of a wide variety of eukaryotic microorganisms by use of a single primer set and holds promise for the development of typing schemes for both T. vaginalis and G. lamblia.

摘要

我们克隆并测序了来自阴道毛滴虫(一种具鞭毛的原生动物寄生虫)的一段可变DNA重复序列。在聚合酶链反应中针对该重复序列进行靶向操作,结果显示,对于多种真核微生物,包括致病性原生动物寄生虫阴道毛滴虫、蓝氏贾第鞭毛虫、杜氏利什曼原虫、三种锥虫以及四种棘阿米巴;非致病性原生动物四膜虫和嗜热栖热四膜虫;以及一种酵母酿酒酵母,都产生了复杂且强烈的产物模式。每种微生物都呈现出独特的重复序列模式。例如,六种临床阴道毛滴虫分离株呈现出一种特征模式。八种蓝氏贾第鞭毛虫分离株呈现出两种特征模式类型中的一种。该方法与人类DNA或来自解脲脲原体和人型支原体等原核生物的DNA均无反应。这种方法可能有助于通过使用单一引物组来检测多种真核微生物,并有望为阴道毛滴虫和蓝氏贾第鞭毛虫的分型方案的开发带来希望。

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