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福尔马林固定粪便中蓝氏贾第鞭毛虫磷酸丙糖异构酶基因的聚合酶链反应扩增

PCR amplification of triosephosphate isomerase gene of Giardia lamblia in formalin-fixed feces.

作者信息

Molina Nora, Polverino Daniela, Minvielle Marta, Basualdo Juan

机构信息

Cátedra de Microbiología y Parasitología, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, Argentina.

出版信息

Rev Latinoam Microbiol. 2007 Jan-Jun;49(1-2):6-11.

Abstract

Giardia lamblia (syn G. intestinalis, G. duodenalis) is the intestinal protozoan producing non-bacterial diarrhea most prevalent in the world. PCR genotype classification of Giardia in feces depends on the quality and quantity of purified DNA and the removal of a great number of inhibitors. The aim of this study was to adapt a PCR protocol to the amplification of the triosephosphate isomerase (tpi) gene of Giardia lamblia in formalin-fixed feces. The tpi gene of G. lamblia was amplified in 28 of the 34 analyzed samples (82.35%) and the B genotype was obtained in all cases. Two major modifications were implemented to improve the performance of PCR from formolated fecal matter. One of these improvements was the use of polyvinylpyrrolidone (PVP) and the other was the addition of bovine serum albumin (BSA). The PCR protocol used in this study showed an amplification percentage exceeding the values reported by other authors with high sensibility and specificity.

摘要

蓝氏贾第鞭毛虫(同义词:肠贾第鞭毛虫、十二指肠贾第鞭毛虫)是世界上引起非细菌性腹泻最常见的肠道原生动物。粪便中贾第鞭毛虫的聚合酶链反应(PCR)基因型分类取决于纯化DNA的质量和数量以及大量抑制剂的去除。本研究的目的是调整一种PCR方案,用于扩增福尔马林固定粪便中蓝氏贾第鞭毛虫的磷酸丙糖异构酶(tpi)基因。在34个分析样本中的28个(82.35%)中扩增出了蓝氏贾第鞭毛虫的tpi基因,且所有病例均获得了B基因型。为提高来自福尔马林固定粪便样本的PCR性能实施了两项主要改进措施。其中一项改进是使用聚乙烯吡咯烷酮(PVP),另一项是添加牛血清白蛋白(BSA)。本研究中使用的PCR方案显示出的扩增百分比超过了其他作者报道的值,具有高敏感性和特异性。

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