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通过线性探针分析和核苷酸测序监测,捷克共和国分离株中对逆转录酶核苷抑制剂耐药的人类免疫缺陷病毒1型毒株

Human immunodeficiency virus 1 strains resistant to nucleoside inhibitors of reverse transcriptase in isolates from the Czech Republic as monitored by line probe assay and nucleotide sequencing.

作者信息

Reinis M, Vandasová J, Stanková M, Linka M, Brůcková M

机构信息

National Reference Laboratory on AIDS, National Institute of Public Health, Prague, Czech Republic.

出版信息

Acta Virol. 2001;45(5-6):279-86.

Abstract

The genetic resistance to nucleoside inhibitors of the reverse transcriptase (RT) of human immunodeficiency virus I (HIV-1) isolates in the Czech Republic was examined by a line probe assay (LiPA) and nucleotide sequencing. The results of LiPA analysis of 294 blood specimens obtained from 156 patients revealed a high incidence of mutations in the RT gene related to resistance to various drugs (67.3%) in various combinations. Mutations in RT gene (M41L, K70R and T215Y/F) conferring the resistance to zidovudine (ZDV) were most frequent (62.6%), that (M184V) responsible for the resistance to lamivudine (3TC) was less frequent (33.7%), while those linked to the resistance to dideoxyinosine (ddl) and dideoxyinosine together with dideoxycytidine (ddl/ddC) were rather rare (6.5% and 5.1%, respectively). LiPA gave a high rate of uninterpretable results due to codon hybridization failure, especially in HIV-1 isolates of non-B subtype. Thirty-two specimens were analyzed also by direct sequencing of a part of RT gene. The results obtained by LiPA and the sequencing were highly concordant for codons successfully analyzed by both methods, but the sequencing provided information also about the codons that could not be analyzed by LiPA. A high prevalence of resistant strains in the Czech Republic and their heterogeneity justifies a regular HIV-1 resistance testing. LiPA turned out as a fast, powerful and most reliable tool for such a purpose. However, due to an increasing diversity of HIV-1 strains circulating in the Czech Republic, LiPA cannot replace the nucleotide sequence analysis.

摘要

采用线性探针分析法(LiPA)和核苷酸测序技术,对捷克共和国人类免疫缺陷病毒I型(HIV-1)分离株逆转录酶(RT)对核苷类抑制剂的遗传耐药性进行了检测。对156例患者的294份血液标本进行LiPA分析的结果显示,RT基因中与多种药物耐药相关的突变发生率很高(67.3%),且存在多种组合形式。赋予齐多夫定(ZDV)耐药性的RT基因突变(M41L、K70R和T215Y/F)最为常见(62.6%),赋予拉米夫定(3TC)耐药性的突变(M184V)较少见(33.7%),而与去羟肌苷(ddI)以及去羟肌苷和双脱氧胞苷联合使用(ddI/ddC)耐药相关的突变则相当罕见(分别为6.5%和5.1%)。由于密码子杂交失败,LiPA得出的无法解读结果的比例很高,尤其是在非B亚型的HIV-1分离株中。还对32份标本进行了RT基因部分片段的直接测序。LiPA和测序法对两种方法均成功分析的密码子得出的结果高度一致,但测序法还提供了LiPA无法分析的密码子的信息。捷克共和国耐药菌株的高流行率及其异质性证明有必要定期进行HIV-1耐药性检测。事实证明,LiPA是实现这一目的的快速、强大且最可靠的工具。然而,由于捷克共和国流行的HIV-1毒株多样性不断增加,LiPA无法取代核苷酸序列分析。

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