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Genotyping of echinococcus granulosus isolates from human clinical samples based on sequencing of mitochondrial genes in Iran, Tehran

机译:基于伊朗德黑兰线粒体基因测序的人类临床样本中细粒棘球菌分离株的基因分型

摘要

Background: The present study was aimed to investigate molecular diversity of Echinococcus granulosus isolates collected from human clinical samples using two mitochondrial genes cox1 and nad1 in Iran. Methods: Forty seven human hydatid cysts were collected through surgery from two hospitals in Tehran during 2010-2012. To determine the fertility of protoscoleces, the cyst fluids were subjected to morphological microscopic examinations. Protoscoleces were removed from each cyst and their total genomic DNAs were extracted. PCR was performed to amplify fragments of 450 and 400 base pair (bp) for cox1 and nad1 genes, respectively. Genotype diversity and sequence variation of the strains were studied by bioinformatics software and in comparison with those mtDNA sequences already deposited in GenBank. Results: Sixteen, (53.3), 13 (43.3), and 1 (3.3) samples were related to lung, liver, and spleen, respectively. The remained 17 unfertile samples were excluded from the study. From the 29 isolates, 86.7 (n=26) and 10 (n=3) were related to G1, and G3 genotypes, respectively. The sole isolate with G6 genotype was obtained from lung sample. Analysis of concatenated sequences of cox1+nad1 indicated the presence of 11 haplotypes among our strains that were related to genotypes G1 (n=9), G3 (n=1) and G6 (n=1). Conclusion: In consistent to other reports from Iran, genotypes G1, G3, and G6 were observed in our human isolates. The rate of G3 genotype was however higher than other studies implying that human can be considered as a new appropriate host for G3 genotype. Further studies with more sample size from different geographic areas of Iran are needed for E. granulosus mapping.
机译:背景:本研究旨在调查在伊朗使用两个线粒体基因cox1和nad1从人类临床样品中收集的细粒棘球oc分离物的分子多样性。方法:在2010-2012年期间,通过手术从德黑兰的两家医院收集了47例人包虫囊肿。为了确定原卵的繁殖力,对囊肿液进行形态学显微镜检查。从每个囊肿中去除原核,并提取其总基因组DNA。进行PCR分别扩增cox1和nad1基因的450和400个碱基对(bp)的片段。通过生物信息学软件研究了菌株的基因型多样性和序列变异,并与已经保存在GenBank中的那些mtDNA序列进行了比较。结果:分别与肺,肝和脾相关的样本有16个,(53.3),13个(43.3)和1个(3.3)。剩余的17个不育样品被排除在研究之外。在这29个菌株中,分别有86.7个(n = 26)和10个(n = 3)与G1和G3基因型有关。从肺样品获得具有G6基因型的唯一分离株。 cox1 + nad1串联序列的分析表明,我们的菌株中存在11个单倍型,它们与基因型G1(n = 9),G3(n = 1)和G6(n = 1)相关。结论:与伊朗的其他报道一致,在我们的人类分离物中观察到基因型G1,G3和G6。然而,G3基因型的发生率高于其他研究,这意味着可以将人类视为G3基因型的新的合适宿主。颗粒大肠杆菌的作图需要对来自伊朗不同地理区域的更多样本量进行进一步研究。

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