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[土耳其分离出的HIV-1毒株的尸检分子流行病学]

[Postmortem Molecular Epidemiology of HIV-1 Strains Isolated in Turkey].

作者信息

Ziyade Nihan, Sayan Murat, Elgörmüş Neval, Büyük Yalçın

机构信息

Council of Forensic Medicine, Postmortem Microbiology Laboratory, Istanbul,Turkey.

Kocaeli University Faculty of Medicine, PCR Unit, Kocaeli, Turkey.

出版信息

Mikrobiyol Bul. 2019 Oct;53(4):374-387. doi: 10.5578/mb.68681.

DOI:10.5578/mb.68681
PMID:31709935
Abstract

Human immunodeficiency virus (HIV) comprises two genotypes, namely HIV-1 (group M, N, O and P) and HIV-2 (group A to H), which differ in their envelope glycoproteins and other antigenic epitopes despite their morphological and biological resemblance.Group M of HIV-1 responsible for 95% of HIV infections worldwide is composed of nine subgroups. In addition to subgroups, group M contains also two recombinant forms, known as circulating recombinant form (CRF) and unique recombinant form (URF). The first case of HIV/acquired immun deficiency virus (AIDS) in Turkey was reported in 1985 and the current number of cases reached a total of 18.557 including 1736 with AIDS based upon the surveillance data of Ministry of Health between October 1985 and November 2018. The aim of this study was to determine the prevalence of HIV-1 strains isolated from HIV positive autopsy cases detected by HIV polymerase chain reaction (PCR) and determine drug resistance. Twenty eight cases [17 males, 11 female: age ranged between 3 months and 66 years (median: 35 years)] found to be HIV positive among the autopsy cases sent for HIV1 PCR study and serological screening between 2011-2017 were recruited in the study. For identification of subtypes in HIV-1 isolates, most-preferred analysis tool was used [HIVdb Stanford University Genotypic Resistance Interpretation Algorithm (www.hivdb.stanford.edu)]. Phylogenetic tree was made according to direct sequencing of HIV-1 reverse transcriptase (pol) region and phylogenetic analysis was evaluated in 23 cases. Los Alamos National Laboratory were trimmed from full-length genomes. Phylogenetic analysis of the 870 base pair of the pol gene region was performed using CLC Sequence Viewer v8.0 (Qiagen Aarhus A/S, www.qiagenbioinformatics.com) software. The phylogenetic tree was obtained according to the neighbor-joining method and the Jukes-Cantor nucleotide distance scale and bootstrap value was set at 1000. In our study, subtype B was found to be most frequent type (39.3%; 11/28). Subtype A (17.9%; 5/28), CRF02_AG (14.3%; 4/28), subtype C (10.7%; 3/28), B+CRF02_AG recombinant (3.6%; 1/28), CRF01_AE (3.6%; 1/28), subtype D (3.6%; 1/28), as well as subtype F (3.6%; 1/28) and subtype G (3.6%; 1/28) strains were also detected in the circulation. Analysis of our results showed that 32.1% (9/28) of the samples exhibited resistance mutations. Detected mutations were as follows: M41L, T215C, K65R, M184V, responsible for nucleoside reverse transcriptase inhibitor (NRTI) resistance; K103N, Y181C, G190A, responsible for non-nucleoside reverse transcriptase inhibitor (NNRTI) resistance; D30N, M46I, responsible for protease inhibitor (PI) resistance. NRTI, NNRTI and PI mutation rates in the samples were found as 21.4%, 7.1% and 3.6%, respectively. Although number of samples analyzed in our study is low, we can propose that they resemble the strains circulating in Turkey. The results of our study; although the subtype B is still dominant in our country, it supports other studies reporting that there are non-B subtypes and an increase in CRF rates in recent years. Phylogenetic analysis is widely regarded as the gold standard technique to determine the subtypes of HIV-1. Molecular epidemiologic studies related to HIV may be important in monitoring HIV subtype patterns and spreading pathways in that country. As a result; the opportunity to collect postmortem HIV sequences in a database appears to have occurred, and as this database expands, its usability is available. Therefore, it is thought that HIV subtypes and mutation information may be useful.

摘要

人类免疫缺陷病毒(HIV)有两种基因型,即HIV-1(M、N、O和P组)和HIV-2(A至H组),尽管它们在形态和生物学特征上相似,但其包膜糖蛋白和其他抗原表位有所不同。导致全球95%HIV感染的HIV-1 M组由9个亚组组成。除亚组外,M组还包含两种重组形式,即循环重组形式(CRF)和独特重组形式(URF)。土耳其于1985年报告了首例HIV/获得性免疫缺陷病毒(AIDS)病例,根据卫生部1985年10月至2018年11月的监测数据,目前病例总数达18557例,其中1736例为艾滋病患者。本研究的目的是确定通过HIV聚合酶链反应(PCR)检测出的HIV阳性尸检病例中分离出的HIV-1毒株的流行情况并确定耐药性。在2011 - 2017年间送去进行HIV1 PCR研究和血清学筛查的尸检病例中,有28例[17例男性,11例女性:年龄在3个月至66岁之间(中位数:35岁)]被发现HIV呈阳性,并被纳入本研究。为鉴定HIV-1分离株中的亚型,使用了最优选的分析工具[斯坦福大学HIVdb基因型耐药性解读算法(www.hivdb.stanford.edu)]。根据HIV-1逆转录酶(pol)区域的直接测序构建系统发育树,并对23例病例进行了系统发育分析。从全长基因组中截取了洛斯阿拉莫斯国家实验室的数据。使用CLC Sequence Viewer v8.0(Qiagen Aarhus A/S,www.qiagenbioinformatics.com)软件对pol基因区域的870个碱基对进行系统发育分析。根据邻接法和Jukes-Cantor核苷酸距离尺度获得系统发育树,自展值设为1000。在我们的研究中,B亚型是最常见的类型(39.3%;11/28)。还检测到了A亚型(17.9%;5/28)、CRF02_AG(14.3%;4/28)、C亚型(10.7%;3/28)、B + CRF02_AG重组型(3.6%;1/28)、CRF01_AE(3.6%;1/28)、D亚型(3.6%;1/28)以及F亚型(3.6%;1/28)和G亚型(3.6%;1/28)毒株在流行中。对我们结果的分析表明,32.1%(9/28)的样本存在耐药突变。检测到的突变如下:导致核苷类逆转录酶抑制剂(NRTI)耐药的M41L、T215C、K65R、M184V;导致非核苷类逆转录酶抑制剂(NNRTI)耐药的K103N、Y181C、G190A;导致蛋白酶抑制剂(PI)耐药的D30N、M46I。样本中的NRTI、NNRTI和PI突变率分别为21.4%、7.1%和3.6%。尽管我们研究中分析的样本数量较少,但我们可以认为它们与土耳其流行的毒株相似。我们研究的结果;尽管B亚型在我国仍然占主导地位,但它支持了其他研究报告,即存在非B亚型且近年来CRF率有所增加。系统发育分析被广泛认为是确定HIV-1亚型的金标准技术。与HIV相关的分子流行病学研究对于监测该国HIV亚型模式和传播途径可能很重要。结果;似乎出现了在数据库中收集死后HIV序列的机会,并且随着这个数据库的扩大,其可用性也具备了。因此,认为HIV亚型和突变信息可能会有用。

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