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目的利用多位点数目可变串联重复序列技术,初步探索甘肃省结核分枝杆菌的基因型及其分布,为防治结核病提供科学依据。方法选择15个VNTR位点,设计引物,采用PCR扩增、琼脂糖凝胶电泳检测,并利用BioNumerics 4.5软件进行DNA指纹图谱多态性分析。结果多位点数目可变串联重复序列(MLVA)检测显示,215株结核分枝杆菌呈现7个基因群127种基因型,分别为a、b、c、d、e、f和g基因群,其中e群74.88%(161/215)为主要流行型(spoligotyping鉴定为北京家族基因型);有抗结核治疗史患者菌株成簇率高于无抗结核治疗史患者,差异有统计学意义(χ2=3.91,P=0.046)。结论兰州地区结核分枝杆菌基因DNA指纹图谱呈现多态性,存在主要流行株,MLVA有助于为当地政府部门制定具体的结核病防治政策和公共卫生应急方案提供科学依据。
Objective To explore the genotypes and distribution of Mycobacterium tuberculosis in Gansu province by using variable number of tandem repeats and provide a scientific basis for the prevention and treatment of tuberculosis. Methods Fifteen VNTR loci were selected, primers were designed, and PCR amplification and agarose gel electrophoresis were performed. The DNA fingerprinting polymorphism was analyzed by BioNumerics 4.5 software. Results A total of 127 genotypes were found in 215 Mycobacterium tuberculosis strains, which were a, b, c, d, e, f and g, respectively. The results of multiple locus variable tandem repeat (MLVA) Among them, 74.88% (161/215) of e group were predominantly epidemic (Beijing family genotype was identified by spoligotyping), and the clustering rate of patients with anti-TB treatment was higher than those without anti-TB treatment (χ2 = 3.91, P = 0.046). Conclusion The genomic DNA fingerprinting of Mycobacterium tuberculosis in Lanzhou is polymorphic and there are major epidemic strains. MLVA can help to provide a scientific basis for local government departments to formulate specific tuberculosis prevention and control policies and public health emergency response plans.