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使用聚合酶链反应-限制性片段长度多态性(RFLP)法测定阴道毛滴虫基因型

Determination of Trichomonas vaginalis Genotypes Using PCR-Restriction Fragment Length Polymorphism (RFLP).

作者信息

Demirağ Serpil, Malatyalı Erdoğan, Ertuğ Sema, Ertabaklar Hatice

机构信息

Adnan Menderes Üniversitesi Tıp Fakültesi, Aile Hekimliği Anabilim Dalı, Aydın, Türkiye.

Adnan Menderes Üniversitesi Tıp Fakültesi, Parazitoloji Anabilim Dalı, Aydın, Türkiye.

出版信息

Turkiye Parazitol Derg. 2017 Dec;41(4):188-191. doi: 10.5152/tpd.2017.5496.

Abstract

OBJECTIVE

Trichomonas vaginalis is an anaerobic protozoon and the most common non-viral sexually transmitted pathogen. The present study was designed to determine the genotypes of T. vaginalis using polymerase chain reaction (PCR)-restriction fragment length polymorphism of actin gene.

METHODS

A total of 20 isolates from symptomatic females isolated and cryopreserved at Adnan Menderes University, Research and Training Hospital Parasitology Laboratory were included. The isolates from liquid nitrogen were thawed and grown in trypticase-yeast extract-maltose medium prior to the study. Following nucleic acid extraction, the actin gene of T. vaginalis was amplified using nested PCR and amplicons were concentrated with phenol-chloroform-isoamyl alcohol precipitation. The final products were digested with HindII, MseI, and RsaI and were visualized using agarose gel electrophoresis.

RESULTS

Most isolates were actin genotype E (n=9, 45%). The remaining isolates were genotype G (n=7, 35%), genotype N (n=1, 5%), and genotype H (n=1, 5%); two were mixed genotypes of E and H (10%).

CONCLUSION

To the best of our knowledge, this study is the first to provide data on T. vaginalis genotypes in Turkey. However, further studies should be conducted to understand the molecular epidemiology of T. vaginalis at the national and global levels.

摘要

目的

阴道毛滴虫是一种厌氧原生动物,也是最常见的非病毒性传播病原体。本研究旨在利用肌动蛋白基因的聚合酶链反应(PCR)-限制性片段长度多态性来确定阴道毛滴虫的基因型。

方法

纳入了阿德南·曼德列斯大学研究与培训医院寄生虫学实验室分离并冷冻保存的20株有症状女性的分离株。在研究前,将液氮中的分离株解冻,并在胰蛋白酶-酵母提取物-麦芽糖培养基中培养。核酸提取后,使用巢式PCR扩增阴道毛滴虫的肌动蛋白基因,扩增产物用酚-氯仿-异戊醇沉淀法浓缩。最终产物用HindII、MseI和RsaI进行酶切,并用琼脂糖凝胶电泳进行可视化分析。

结果

大多数分离株为肌动蛋白基因型E(n = 9,45%)。其余分离株为基因型G(n = 7,35%)、基因型N(n = 1,5%)和基因型H(n = 1,5%);两株为E和H的混合基因型(10%)。

结论

据我们所知,本研究首次提供了土耳其阴道毛滴虫基因型的数据。然而,应开展进一步研究以了解国家和全球层面阴道毛滴虫的分子流行病学。

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