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白俄罗斯结核病患者中耐异烟肼结核分枝杆菌分离株的突变检测

Detection of mutation in isoniazid-resistant Mycobacterium tuberculosis isolates from tuberculosis patients in Belarus.

作者信息

Bostanabad S Z, Titov L P, Bahrmand A, Nojoumi S A

机构信息

Belarusian Research Institute for Epidemiology and Microbiology, Filimonova 23, Minsk, Belarus.

出版信息

Indian J Med Microbiol. 2008 Apr-Jun;26(2):143-7. doi: 10.4103/0255-0857.40528.

Abstract

The aim of this study was to investigate the frequency, location and type of katG mutations in Mycobacterium tuberculosis strains isolated from patients in Belarus. Forty two isoniazid-resistant isolates were identified from sputum of 163 patients with active pulmonary tuberculosis. Drug susceptibility testing was determined by using CDC standard conventional proportional method and BACTEC system. Standard PCR method for detection of isoniazid resistance associated mutations was performed by katG gene amplification and DNA sequencing. Most mutations were found in katG gene codons 315, 316 and 309. Four types of mutations were identified in codon 315: AGC-->ACC (n=36) 85%, AGC-->AGG (n=1) 2.3%, AGC-->AAC (n=2) 4.7%, AGC-->GGC (n=1) 2.3%. One type of mutation was found in codon 316: GGC-->AGC (n=18) 41.4%, four types of mutations were detected in codon 309: GGT-->GGT (n=7) 16.1%, GGT-->GCT (n=4) 9.2%, GGT-->GTC (n=3)6.9%, GGT-->GGG (n=1) 2.7%. The highest frequency of mutations sharing between primary and secondary infections was found in codon 315.

摘要

本研究的目的是调查从白俄罗斯患者中分离出的结核分枝杆菌菌株中katG基因突变的频率、位置和类型。从163例活动性肺结核患者的痰液中鉴定出42株耐异烟肼菌株。采用美国疾病控制与预防中心(CDC)标准常规比例法和BACTEC系统进行药敏试验。通过katG基因扩增和DNA测序,采用标准PCR方法检测与异烟肼耐药相关的突变。大多数突变发生在katG基因的315、316和309密码子。在315密码子中鉴定出四种突变类型:AGC→ACC(n = 36)85%,AGC→AGG(n = 1)2.3%,AGC→AAC(n = 2)4.7%,AGC→GGC(n = 1)2.3%。在316密码子中发现一种突变类型:GGC→AGC(n = 18)41.4%,在309密码子中检测到四种突变类型:GGT→GGT(n = 7)16.1%,GGT→GCT(n = 4)9.2%,GGT→GTC(n = 3)6.9%,GGT→GGG(n = 1)2.7%。在315密码子中,原发感染和继发感染之间共享的突变频率最高。

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