沧州市结核分枝杆菌间隔区寡核苷酸分型研究

摘要

目的：初步了解沧州市结核分枝杆菌临床分离株的基因多态性和分子流行病学特征，探讨各基因型与表型耐药之间的关系。方法收集结核病患者经临床分离培养得到的结核分枝杆菌菌株及相应病例信息，提取菌株DNA，采用间隔区寡核苷酸基因分型法进行基因分型研究，采用BACTECTMMGITTM960液体培养技术进行一线抗结核药物敏感试验。运用BioNumerics 5.0软件进行聚类分析，运用Graphpad Prism 5.0软件进行数据分析。结果154例患者中男109例，女45例；初治121例，复治33例；34例有吸烟史，12例合并糖尿病。48株（31.2%）至少对链霉素、异烟肼、利福平、乙胺丁醇中的1种药物耐药。复治患者的耐药率（63.6%）显著高于初治患者耐药率（22.3%）。典型北京型菌株占所有菌株的91.6%，非典型北京型菌株占8.4%。典型北京型与非典型北京型在患者性别、治疗情况、吸烟史、糖尿病史及耐药分布上的差异均无统计学意义。结论沧州市结核分枝杆菌呈现明显的基因多态性。典型北京型菌株是该地区的优势菌株，且基因型与耐药性无关。%Objective To identify the genetic polymorphism and molecular epidemiological characteristics of Mycobacterium tuberculosis (TB) clinical isolates in Cangzhou, and explore the association between genotypes and drug resistance phenotypes. Methods Mycobacterium tuberculosis isolates were collected from TB patients treated in the Cangzhou Infectious Disease Hospital, and corresponding clinical data were also collected. The genomic DNA was extracted. Spacer oligonucleotide typing (spoligotyping) was applied to genotype the isolates. Drug susceptibility testing of first-line anti-TB drugs (streptomycin, isoniazid, rifampin, ethambutol) was performed using the BACTECTMMGITTM960 liquid medium. Cluster analysis was done by BioNumerics 5.0. Data was analyzed by Graphpad Prism 5.0 software. Results Of the 154 patients, 109 were male and 45 were female. And 121 patients were treated for the first time, 33 were retreatment, 34 had smoking history, 12 were complicated with diabetes mellitus. Fourty-eight strains (31.2%) were resistant to at least one of streptomycin, inoniazid, rifampicin and ethambutol. Initial drug resistance rate was 22.3%and acquired drug resistance rate was 63.6%. The drug resistance rate was significantly higher in retreatment patients than that of new cases. The Beijing family strain accounted for 91.6% of all isolates and the non-Beijing family strain accounted for 8.4%. There were no significant differences in gender, treatment history, smoking history and diabetes history between patients with the Beijing genotype and patients with non-Beijing genotype. Conclusion Mycobacterium tuberculosis in Cangzhou exhibits high genetic diversity. The Beijing genotype is the predominant no association with drug resistance.