福尔马林固定粪便中兰氏贾第虫三磷酸异构酶基因的PCR扩增。

Nora Molina, Daniela Polverino, Marta Minvielle, Juan Basualdo
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引用次数: 0

摘要

蓝氏贾第鞭毛虫是世界上最常见的引起非细菌性腹泻的肠道原生动物。粪便中贾第鞭毛虫的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。本研究的目的是采用PCR方法在固定福尔马林的粪便中扩增兰氏贾第鞭毛虫三磷酸异构酶(tpi)基因。34份样本中有28份(82.35%)扩增到兰氏螺旋体tpi基因,全部为B基因型。为了提高从粪便中提取的PCR的性能,我们进行了两个主要的改进。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)。本研究使用的PCR方案显示扩增率超过其他作者报道的值,具有较高的敏感性和特异性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
PCR amplification of triosephosphate isomerase gene of Giardia lamblia in formalin-fixed feces.

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.

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