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利用聚合酶链反应-限制性片段长度多态性技术测定伊朗西南部舒什县屠宰牲畜的基因型

Determination of Genotypes in Livestock Slaughtered in Shush County, Southwest Iran Using PCR-RFLP.

作者信息

Fallahizadeh S, Arjmand R, Jelowdar A, Rafiei A, Kazemi F

机构信息

Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

Student Research Committee, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

出版信息

Helminthologia. 2019 Sep 1;56(3):196-201. doi: 10.2478/helm-2019-0023. eCollection 2019 Sep.

Abstract

Echinococosis is a zoonotic disease caused by the larval stages of spp. that occurs in most parts of the world. Herein, we aimed to evaluate the genotypes of isolated hydatid cysts from slaughtered animals in Shush county, southwestern Iran. Totally, 96 hydatid cysts were collected, including 11 buffaloes, 13 cattle, 12 goat and 60 sheep. The PCR was done by a primer pair (BDI and 4s) to amplify ITS1 fragment. Four restriction endonucleases including AluI, HpaII, RsaI, and TaqI were used for RFLP products and enzymatic reactions were electrophoresed. Finally, twenty PCR products were sent for sequencing and phylogenetic tree was drawn with MEGA6. Molecular identification of 96 hydatid cysts demonstrated a distinctive 1000 bp fragment in all samples from four animal hosts. RFLP analysis showed similar digestion patterns in all samples. AluI digestion yielded 800 bp and 200 bp fragments, HpaII digestion made 700 bp and 300 bp fragments and RsaI digestion entailed 655 and 345segments. Moreover, TaqI rendered no digestion pattern on rDNA-ITS1 region. Additionally, sensu stricto (G1-3 complex) was the prevailing genotype in all livestock samples, according to PCR-RFLP and sequencing analyses.

摘要

棘球蚴病是一种由棘球绦虫属幼虫阶段引起的人畜共患病,在世界大部分地区都有发生。在此,我们旨在评估从伊朗西南部舒什县屠宰动物中分离出的包虫囊肿的基因型。总共收集了96个包虫囊肿,包括11头水牛、13头牛、12只山羊和60只绵羊。通过一对引物(BDI和4s)进行PCR扩增ITS1片段。使用包括AluI、HpaII、RsaI和TaqI在内的四种限制性内切酶对RFLP产物进行酶切反应,并进行电泳。最后,将20个PCR产物送去测序,并用MEGA6绘制系统发育树。对96个包虫囊肿的分子鉴定表明,来自四种动物宿主的所有样本中都有一个独特的1000 bp片段。RFLP分析显示所有样本的消化模式相似。AluI酶切产生800 bp和200 bp片段,HpaII酶切产生700 bp和300 bp片段,RsaI酶切产生655和345片段。此外,TaqI在rDNA-ITS1区域没有产生消化模式。另外,根据PCR-RFLP和测序分析,狭义细粒棘球绦虫(G1-3复合体)是所有家畜样本中的主要基因型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c0e4/6799581/127ff1fb5ac5/helm-56-196-g001.jpg

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