Mondragón-Barreto M, Vázquez-Chacón C A, Barrón-Rivero C, Acosta-Blanco P, Jost K C, Balandrano S, Olivera-Díaz H
Departamento de Biología Molecular y Departamento de Micobacterias, Instituto de Diagnóstico y Referencia Epidemiológicos, México, D.F., México.
Salud Publica Mex. 2000 Nov-Dec;42(6):484-9. doi: 10.1590/s0036-36342000000600003.
To compare three methods: Biochemical tests, high-performance liquid chromatography (HPLC) and polymerase chain reaction-restriction fragments length polymorphism (PCR-RFLP), for the identification of mycobacteria, and to perform a cost-benefit analysis to define an optimum identification algorithm.
One-hundred-and-seven mycobacteria isolates were identified by the three methods at Instituto de Diagnóstico y Referencia Epidemiológicos, between February of 1999 and January of 2000 and the results were compared with those of a reference laboratory using the Q-Cochran statistical test.
PCR-RFLP was the most rapid and specific procedure but also the most expensive; biochemical tests excelled for identification of Mycobacterium tuberculosis, but were lengthy and expensive for other mycobacteria; HPLC ranked in the middle for price, speed and specificity.
Considering the expected proportion of M. tuberculosis, the following algorithm was proposed: Initially, biochemical tests should be performed; if the results indicate a non-tuberculous mycobacteria, the isolate should be analyzed with HPLC; if results are unclear, the isolate should be analyzed using PCR-RFLP. Isolates showing a previously undescribed PCR-RFLP pattern should be characterized by DNA sequencing.
比较三种用于鉴定分枝杆菌的方法:生化试验、高效液相色谱法(HPLC)和聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP),并进行成本效益分析以确定最佳鉴定算法。
1999年2月至2000年1月期间,在流行病学诊断与参考研究所,用这三种方法对107株分枝杆菌分离株进行鉴定,并使用Q- Cochr an统计检验将结果与参考实验室的结果进行比较。
PCR-RFLP是最快速、特异的方法,但也是最昂贵的;生化试验在结核分枝杆菌鉴定方面表现出色,但对其他分枝杆菌鉴定耗时且昂贵;HPLC在价格、速度和特异性方面排名居中。
考虑到结核分枝杆菌的预期比例,提出以下算法:首先进行生化试验;如果结果表明为非结核分枝杆菌,则用HPLC分析该分离株;如果结果不明确,则用PCR-RFLP分析该分离株。显示出以前未描述的PCR-RFLP模式的分离株应以DNA测序进行鉴定。