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伊朗西阿塞拜疆地区采用PCR-RFLP法对动物分离的包虫囊肿标本中细粒棘球绦虫菌株的鉴定

Identification of Echinococcus granulosus Strains in Isolated Hydatid Cyst Specimens from Animals by PCR-RFLP Method in West Azerbaijan - Iran.

作者信息

Hanifian Haleh, Diba Kambiz, Tappeh Khosrow Hazrati, Mohammadzadeh Habib, Mahmoudlou Rahim

机构信息

Dept. of Medical Parasitology and Mycology, Faculty of medicine, Urmia University of Medical Sciences, Urmia, Iran.

Cellular and Molecular Research center, Urmia University of Medical Seiences, Urmia-Iran.

出版信息

Iran J Parasitol. 2013 Jul;8(3):376-81.

Abstract

BACKGROUND

The aim of this study was DNA extraction from protoscolecses of Echinococcus granulosus and identification of these strains in West-Azerbaijan Province, north western Iran.

METHODS

Thirty one livestock isolates from sheep and cattle were collected from abattoirs of the province. To investigate the genetic variation of the isolates, after DNA extraction by Glass beads-phenol chloroform method; PCR-RLFP analysis of rDNA-ITS1 was performed using three different restriction enzymes of Taq 1, Rsa 1 and Alu 1.

RESULT

Amplified PCR products for all isolates were 1000bp band which is expected band in sheep strains (G1-G3 complex). The results of RFLP analysis also were the same for all isolates. PCR-RFLP patterns restriction enzymes were identical as follows, Rsa1 bands under UV showed two bands approximately 655bp and 345bp. Alu1 bands were as follows: two approximately 800bp and 200bp and Taq1 did not cut any region and bands were approximately 1000 bp in all samples.

CONCLUSIONS

Based on PCR-RFLP patterns of ITS1 fragment produced with endonucleases enzyme digestion in animal isolates, it can be concluded that a single strain of E. granulosus (sheep strain or G1-G3 complex) is dominant genotype in this province.

摘要

背景

本研究的目的是从细粒棘球绦虫原头节中提取DNA,并对伊朗西北部西阿塞拜疆省的这些菌株进行鉴定。

方法

从该省的屠宰场收集了31份来自绵羊和牛的家畜分离株。为了研究分离株的遗传变异,采用玻璃珠-酚氯仿法提取DNA后,使用Taq 1、Rsa 1和Alu 1三种不同的限制性内切酶对rDNA-ITS1进行PCR-RLFP分析。

结果

所有分离株的PCR扩增产物均为1000bp条带,这是绵羊株(G1-G3复合体)的预期条带。RFLP分析结果对所有分离株也相同。PCR-RFLP模式的限制性内切酶相同,如下所示:紫外线下Rsa1条带显示两条条带,约655bp和345bp。Alu1条带如下:两条约800bp和200bp,Taq1未切割任何区域,所有样品中的条带约为1000bp。

结论

根据动物分离株中内切酶消化产生的ITS1片段的PCR-RFLP模式,可以得出结论,细粒棘球绦虫的单一菌株(绵羊株或G1-G3复合体)是该省的主要基因型。

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