Takewaki S, Okuzumi K, Ishiko H, Nakahara K, Ohkubo A, Nagai R
Department of Laboratory Medicine, Faculty of Medicine, University of Tokyo, Japan.
J Clin Microbiol. 1993 Feb;31(2):446-50. doi: 10.1128/jcm.31.2.446-450.1993.
Identification of tuberculous and nontuberculous mycobacteria by biochemical methods is a long-term process that takes up to 8 weeks for completion and requires expertise to interpret the results. In order to detect and differentiate the major pathogenic mycobacterial species, we developed genus-specific primers that amplify the dnaJ gene from the broad spectrum of mycobacterial species and determined the nucleotide sequences within the dnaJ genes from 19 mycobacterial species (Mycobacterium tuberculosis, M. bovis, M. bovis BCG, M. africanum, M. microti, M. kansasii, M. marinum, M. gastri, M. simiae, M. scrofulaceum, M. szulgai, M. gordonae, M. avium, M. intracellulare, M. xenopi, M. fortuitum, M. chelonei, M. haemophilum, and M. paratuberculosis). On the basis of the dnaJ gene sequences, we developed dot blot hybridization analysis with species-specific oligonucleotide probes for the M. tuberculosis complex. M. avium, M. intracellulare, and M. kansaii, allowing a rapid identification of these species following polymerase chain reaction for the dnaJ gene. We conclude that polymerase chain reaction with the genus-specific primer that amplifies the dnaJ genes and subsequent dot blot analysis with species-specific oligonucleotide probes are most useful for differential diagnosis of tuberculosis and nontuberculous mycobacterial infections.
通过生化方法鉴定结核分枝杆菌和非结核分枝杆菌是一个长期过程，完成该过程需要长达8周的时间，并且需要专业知识来解释结果。为了检测和区分主要的致病性分枝杆菌种类，我们开发了属特异性引物，该引物可从广泛的分枝杆菌种类中扩增dnaJ基因，并测定了19种分枝杆菌（结核分枝杆菌、牛分枝杆菌、卡介苗、非洲分枝杆菌、田鼠分枝杆菌、堪萨斯分枝杆菌、海分枝杆菌、胃分枝杆菌、猿分枝杆菌、瘰疬分枝杆菌、苏尔加分枝杆菌、戈登分枝杆菌、鸟分枝杆菌、胞内分枝杆菌、偶发分枝杆菌、龟分枝杆菌、嗜血性分枝杆菌和副结核分枝杆菌）dnaJ基因内的核苷酸序列。基于dnaJ基因序列，我们开发了针对结核分枝杆菌复合群、鸟分枝杆菌、胞内分枝杆菌和堪萨斯分枝杆菌的种特异性寡核苷酸探针斑点杂交分析方法，从而在对dnaJ基因进行聚合酶链反应后能够快速鉴定这些菌种。我们得出结论，使用扩增dnaJ基因的属特异性引物进行聚合酶链反应以及随后使用种特异性寡核苷酸探针进行斑点杂交分析对于结核病和非结核分枝杆菌感染的鉴别诊断最为有用。
