镇江地区结核分枝杆菌DNA指纹图谱分型及耐药性分析

目的：分析镇江地区结核分枝杆菌的DNA指纹图谱分型及耐药情况。方法收集174例疑似结核病患者痰液标本，通过鉴别培养基对硝基苯甲酸（ PNB）和噻吩－2－羟酸肼（ TCH）进行分枝杆菌菌型初筛，再通过本室建立的PCR－ELISA法明确鉴定结核杆菌菌种，用比例法进行药敏实验，用DNA随机扩增多态性（ RAPD）指纹分型法，对其中44株耐药人型结核分枝杆菌进行DNA指纹图谱分型。结果从174份标本中分离出人型结核分枝杆菌141株（81．03％），牛型结核分枝杆菌20株（11．49％），非结核分枝杆菌13株（7．47％），总耐药率42．53％；耐药率由高到低依次为异烟肼（INH）＞氧氟沙星（OFX）＞利福平（RFP）＞链霉素（SM）＞乙胺丁醇（EMB）＞卡那霉素（KM）＞卷曲霉素（ CPM）＞阿米卡星（ AKM）；非结核分枝杆菌对EMB、RFP、SM、INH的耐药率显著高于结核分枝杆菌（ P＜0．01）。复治组对EMB、RFP、SM、INH的耐药率显著高于初治组的耐药率（P＜0．01）。中、老年患者耐药率明显高于青年患者（P＜0．05或P＜0．01）。将44株耐药人型结核杆菌，用RAPD－PCR分型，并用NTsys 2．10e软件作聚类分析，共分为5型（Ⅰ型-Ⅴ型），其中以Ⅰ-Ⅲ型为主，占84．09％。各型的耐药率差异无统计学意义（P＞0．05）。结论本地区肺结核以人型结核分枝杆菌为主，结核杆菌总体耐药率和耐多药率相对较高。 RAPD指纹分型结果显示，该地区结核分枝杆菌具有明显的基因多态性，成簇类型是主要流行菌株。各型与耐药性无明显相关性。

Analysis of DNA fingerprint classification and drug resistance of mycobacterium tuberculosis in Zhenjiang

Objective To analyse DNA fingerprint classification and the situation of drug resistance of mycobacterium tuberculosis in Zhenjiang.Methods Collected 174 cases of suspected TB patients sputum samples,and done early screening to identify bacteria mycobacterium type by differential medium of p-nitrobenzoic acid（ PNB） and thiophene-2-hydroxy acid hy-drazide（TCH）.And then mycobacterium tuberculosis was detected by PCR-ELISA.The susceptibility testing was conducted using the proportional method by means of randomly amplified polymorphic DNA（ RAPD） fingerprinting classification method to analyse DNA fingerprint classification of 44 strains of drug resistanted mycobacterium tuberculosis.Results Among the 174 strains in the study,mycobacterium tuberculosis in mycobacterium hominis 141 strains（81.03%） ,mycobacterium bovis strain 20（11.49%）,13 strains of non mycobacterium tuberculosis（7.47%）,the total drug resistance rate was 42.53%.Drug resist-ance rate from high to low was INH〉OFX〉RFP〉SM〉EMB〉KM〉CPM〉AKM.Non mycobacterium tuberculosis resistant to EMB,RFP,SM,INH were significantly higher than those of mycobacterium tuberculosis.The retreatment group resistance to EMB,RFP,SM,INH were significantly higher than the initial drug resistance rate of the treatment group.In elderly patients, drug resistance rate was higher than that in young patients.44 strains of drug resistanted mycobacterium tuberculosis were di-vided into 5 types by RAPD-PCR classification method,and clustering analysis with NTsys 2.10e software.Type Ⅰ,type Ⅱand typeⅢ are the main types which are accounting for 84.09%.Conclusion Mycobacterium tuberculosis in Zhenjiang mainly is mycobacterium hominis.The prevalence of drug resistance tuberculosis in Zhenjiang was relatively high.There is ob-vious genetic polymorphism in the mycobacterium tuberculosis in Zhenjiang area by RAPD-PCR.And the clustering type was perhaps the main epidemic strains.There was no significant correlation between genotype and drug resestance.