基因芯片法用于检测长沙地区结核分枝杆菌耐药性研究

Identification of drug-resistance gene type in Mycobacterium tuberculosis by gene chip in Hunan province

目的了解长沙市中心医院2013年1月1日至2014年9月30日临床分离的1 031株结核分枝杆菌对利福平和异烟肼的耐药性。方法将结核分枝杆菌培养阳性,并经鉴定为结核分枝杆菌的1 031株菌株,采用绝对浓度法进行常规药敏检测,同时应用基因芯片技术检测利福平和异烟肼耐药相关的rpoB、katG和inhA基因上的位点突变,并对2种结果进行比较。结果基因芯片法检测利福平耐药性中,其敏感株896株,耐药株135株;检测异烟肼耐药性为敏感株901株,耐药株130株。与绝对浓度法比较,基因芯片检测利福平耐药结果与其一致的菌株共1 011株(包含敏感株894株,耐药株117株),符合率为98.00%;基因芯片检测异烟肼耐药结果与其一致的菌株共1 005株(包含敏感株890株,耐药株115株),符合率为97.48%。与利福平耐药相关的rpoB基因的突变位点最常见的为531TCG→TTG突变,占耐药株的51.11%;其次为526CAC→TAC突变耐药株的10.37%;526CAC→GAC共11株菌株突变,占耐药菌株的8.15%。异烟肼耐药主要是由katG315AGC→ACC位突变引起,占耐药株的83.85%,inhA-15C→T突变仅占12.30%。结论基因芯片法与绝对浓度法结果高度一致,可成为筛查利福平和异烟肼耐药性的快速有效方法,长沙地区结核分枝杆菌利福平和异烟肼耐药以rpoB531,526及katG315突变位点为主。

Objective To investigate the drug resistance of 1 031 Mycobacterium tuberculosis to rifampicin and isoniazide in the Center Hospital of Changsha from January 1 ,2013 to September 30 ,2014 .Methods A total of 1 031 strains with positive culture result and identified as strains of Mycobacterium tuberculosis were used absolute concentration method to do the conventional drug susceptibility ,and detected rifampicin and isoniazide resistance gene including rpoB ,katG and inhA gene locus mutation by chip technology ,the results of two methods were compared using card square test statistics .Results By gene chip method ,the sensitive strain of rifampicin was 896 ,the drug-resistant strains was 135 ,the sensitive strains of isoniazide was 901 strains ,130 drug-resistant strains .Compared with the absolute concentration method ,resistance chip detection results were consistent with rifampicin resistant strains 1 011 strains(including 894 drug-resistant strains ,and 117 sensitive strains) ,the coincidence rate was 98 .00% ,consistent with isoniazideresistant strains 1 005 strains(including 890 drug-resistant strains ,115 sensitive strains) ,the coincidence rate was 97 .48% .The most common spot of rifampin resistance related mutations of rpoB gene was 531TCG to TTG ,accounted for 51 .11% ,followed by 526CAC→TAC ,accounted for 10 .37% ,11 strains with 526TCG to TTG ,accounted for 8 .15% .Isoniazid re-sistance was caused by mutations in katG315AGC→ACC resistant strains ,accounted for 83 .85% ,inhA-15C→T mutations accoun-ted for only 12 .30% .Conclusion The results of gene chip method is highly consistent with that of absolute concentration method , could be a fast and effective method for screening rifampicin and isoniazide ,the resistant gene of Mycobacterium tuberculosis to rif-ampicin and isoniazide almost mutate in rpoB531 ,526 and katG315 in Changsha .