某高级中学结核病爆发分离菌株的实验室鉴定

摘要/Abstract

摘要： 目的鉴定从某中学结核病爆发分离菌株的种属。方法对2006年5—8月，辽宁省某高级中学发生的10例痰检阳性病例中的3份痰标本分离的菌株经表型鉴定方法；传统生化方法；扩增hupB基因、dnaA-dnaN 和NTF-1区；Spoligotyping 以及MIRU基因分型；16S rRNA基因、16S-23S ITS和hsp65基因测序以及hsp65基因限制性酶切分析进行鉴定。结果临床分离株经生化方法初步鉴定1份为结核分枝杆菌复合群和2份为非结核分枝杆菌，随后经表型鉴定和扩增hupB基因、dnaA-dnaN 和NTF-1区；Spoligotyping 以及MIRU基因分型，说明属于结核分枝杆菌复合群的菌株为结核分枝杆菌北京基因型现代株，MIRU基因型为223325173533。经16S rRNA基因、16S-23S ITS和hsp65基因测序以及hsp65基因限制性酶切分析说明属于非结核分枝杆菌的菌株为猪分枝杆菌。结论本次从结核病爆发累及病人的标本中分离出结核分枝杆菌北京基因型现代株和猪分枝杆菌，猪分枝杆菌在暴发流行中的意义尚需进一步研究。

关键词: 结核/流行病学, 疾病暴发流行, 分枝杆菌, 结核, 分枝杆菌，非典型性, 细菌学技术

Abstract: ObjectiveTo identify the bacterial species and strain types of isolates from tuberculosis outbreak in a high school. MethodsFrom May to August 2006, one teacher and 64 students developed clinical symptoms of active tuberculosis in a high school, Liaoning Province. Clinical isolates from sputa were collected and identified by phenotypic tests; conventional biochemical methods; amplification of hupB gene, dnaA-dnaN region, NTF-1 region, Spoligotyping and micobacterial interspersed repetitive unit (MIRU) typing; partial sequencing of 16S rRNA, 16S-23S internal transcribed spacer (ITS) region, hsp65 gene (441 bp) and PCR restriction enzyme analysis of the hsp65 gene. ResultsClinical isolates were identified as Mycobacterium tuberculosis complex and non-tuberculous mycobacteria (NTM) by conventional biochemical methods. Subsequent analyzed by phenotypic tests and amplification of hupB gene, dnaA-dnaN region, NTF-1 region, Spoligotyping and MIRU genotyping, these Results indicated that the species identification of mycobacterium tuberculosis complex as mycobacterium tuberculosis Beijing genotype modern strain, the genotype profile of MIRU was 223325173533. By partial sequencing of 16S rRNA, 16S-23S ITS region, hsp65 gene (441 bp) and PCR restriction enzyme analysis of the hsp65 gene supported the species identification of MOTT as M. porcinum. ConclusionThe mycobacterium species isolated from patients involved in tuberculosis outbreak are mycobacterium tuberculosis Beijing genotype modern strain and M. porcinum, the significance of M.porcinum in tuberculosis outbreak should be further studied.

Key words: tuberculosis/epidemiology, disease outbreaks, Mycobacterium tuberculosis, mycobacteria, atypical, bacteriological techniques

王胜芬,王莉,杨立军,姜广路,夏辉,周杨,尚美,李强,赵冰,赵雁林. 某高级中学结核病爆发分离菌株的实验室鉴定[J]. 中国防痨杂志, 2009, 31(10): 573-578.

Wang Shengfen¹, Wang Li², Yang Lijun²,Jiang Guanglu¹, Xia Hui¹, Zhou Yang¹, Shang Mei, Li Qiang¹, Zhao Bing¹, Zhao Yanlin¹. Identification of strains isolated from tuberculosis outbreak in a high school[J]. Chinese Journal of Antituberculosis, 2009, 31(10): 573-578.

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