摘要
为了了解我国结核分枝杆菌耐异烟肼 (INH)分离株katG基因突变情况 ,研究其临床意义。本研究通过聚合酶链反应 (PCR)、PCR 单链构象多态性 (SSCP)和PCR直接测序法 (directsequencing ,DS)分析 92株结核分枝杆菌临床分离株的katG基因。以H37Rv标准株为对照 ,2 3株药物敏感株中 ,2 1株katG基因SSCP分析正常 ,2株异常。 36株耐非INH药物的分离株中 ,2 0株katG基因SSCP正常 ,16株异常。 33株耐INH分离株中 ,高度耐INH的 17株 ,其中 4株为katG基因缺失 ,5株katGSSCP异常 ,8株正常 ;低度耐INH的 16株 ,其中 13株KatG基因SSCP异常 ,3株正常。上述所有katGSSCP异常的分离株DS分析均为 315位密码子突变 ,除 1株为AGC→AAC突变外 ,其余均为AGC→ACC突变。结果表明katG基因突变是结核分枝杆菌耐INH产生的主要分子机制 ,其最常见的 315位密码子突变可引起结核分枝杆菌轻度或中度耐INH。
To understand the mutations of katG gene in M.tuberculosis isolates,and to evaluate their clinical value.Analyzing the katG genes in 92 M.tuberculosis clinical isolates with PCR,PCR SSCP and PCR DS.Mycobacterium tuberculosis strain H37Rv was used as a control. Of 23 drug sensitive isolates,2 displayed abnormal katG SSCP profile.Of 36 non isoniazid resistant isolates,16 had abnormal katG SSCP profile.Of 33 isoniazid resistant isolates,17 were resistant highly to isoniazid,in which 4 katG genes were not amplified and 5 displayed abnormal katG SSCP profile;16 were resistant lowly to isoniazid,in which 13 exhibited abnormal katG SSCP profile.All isolates with abnormal katG SSCP profile had point mutations at codon 315,in which one was AGC→ACC mutation,the others were AGC→AAC mutations .The mutations of katG gene were the most important isoniazid resistant molecular mechanisms in M.tuberculosis.The most common mutations at codon 315 induced the lower MIC for isoniazid.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2000年第1期9-11,共3页
Chinese Journal of Clinical Laboratory Science
基金
解放军总后勤部卫生部科研基金! 970 7
关键词
结核分枝杆菌
DNA序列分析
耐药性
PCR
M.tuberculosis
drug resistance
polymerase chain reaction
direct sequencing
