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利用聚合酶链反应-限制性片段长度多态性技术对伊朗西部伊拉姆省从人和家畜体内分离出的包虫囊肿进行基因分型

Genotyping of Hydatid Cyst Isolated from Human and Domestic Animals in Ilam Province, Western Iran Using PCR-RFLP.

作者信息

Dousti M, Abdi J, Bakhtiyari S, Mohebali M, Mirhendi Sh, Rokni Mb

机构信息

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Iran J Parasitol. 2013 Jan;8(1):47-52.

Abstract

BACKGROUND

Hydatidosis or cystic hydatid disease is one of the most important diseases in human and animals. Identification of strains is important for improvement of control and prevention of disease. The aim of this study was to determine the strains isolated from human and domestic animals in Ilam Province, Iran, using PCR-RFLP method.

METHODS

Respectively, 30 and 4 animal and human hydatid cysts were collected from different slaughterhouses and hospitals of the province. Protoscolices were separated and their DNA genome was extracted by extraction kit. rDNA-ITS1 of each isolated samples was duplicated by BD1(Forward) and 4s (Reverse) Primers. PCR products were studied by electrophoresis and then were digested using TaqI, HpaII, RsaI and AluI restriction enzymes. RFLP products were studied using electrophoresis on 1% agar gel.

RESULT

A fragment of 1000bp was produced from amplification of rDNA-ITS1 of protoscolices using PCR method. After digestion of PCR product by AluI enzyme, 200bp and 800bp, by RsaI, 655bp and 345bp and by HpaII 700bp and 300bp sizes were obtained. TaqI enzyme had no change in fragment size and it remained 1000bp. Considering the method, Ilam strains was specified as E. granulosus sensu stricto (G1-G3).

CONCLUSIONS

Although sheep strain (G1) is dominated in human and different animal in Iran and the world, but more efforts should be done to clarify the true genotype of Ilam strains specified as E. granulosus sensu stricto (G1-G3).

摘要

背景

包虫病或囊性包虫病是人和动物最重要的疾病之一。菌株鉴定对于改善疾病的控制和预防至关重要。本研究的目的是使用PCR-RFLP方法确定从伊朗伊拉姆省的人和家畜中分离出的菌株。

方法

分别从该省不同的屠宰场和医院收集了30个动物包虫囊肿和4个人包虫囊肿。分离出原头蚴,并用提取试剂盒提取其DNA基因组。每个分离样本的rDNA-ITS1用BD1(正向)和4s(反向)引物进行扩增。PCR产物通过电泳进行研究,然后用TaqI、HpaII、RsaI和AluI限制性内切酶进行消化。RFLP产物在1%琼脂凝胶上通过电泳进行研究。

结果

使用PCR方法扩增原头蚴的rDNA-ITS1产生了1000bp的片段。用AluI酶消化PCR产物后,得到200bp和800bp的片段,用RsaI酶消化得到655bp和345bp的片段,用HpaII酶消化得到700bp和300bp的片段。TaqI酶处理后片段大小没有变化,仍为1000bp。根据该方法,伊拉姆菌株被确定为狭义细粒棘球绦虫(G1-G3)。

结论

尽管绵羊株(G1)在伊朗和世界的人和不同动物中占主导地位,但仍应做出更多努力来明确被确定为狭义细粒棘球绦虫(G1-G3)的伊拉姆菌株的真实基因型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a07d/3655239/17a8900f1284/IJPA-8-047-g001.jpg

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