Castro Claudia Marcela, Puerto Gloria, García Luz Mary, Orjuela Dora Leticia, Polo Claudia Llerena, Garzón María Consuelo, Ribón Wellman
Grupo de Micobacterias, Instituto Nacional de Salud, Bogotá, DC, Colombia.
Biomedica. 2007 Sep;27(3):439-46.
Nontuberculous mycobacteria can be saprophytic, pathogenic or opportunistic. The most common diseases produced by these microorganisms are the post-surgical infections due to anesthetic procedures, infections associated with catheters, disseminated cutaneous diseases and pulmonary and central nervous system diseases that especially affect HIV patients. Identification of the nontuberculous mycobacteria can take several weeks and even then, differentiation of complex members is not possible.
The PCR-restriction analysis (PRA) technique was evaluated as a method for genotypic identification of nontuberculous mycobacteria isolated of clinical samples located in the culture collection of the Instituto Nacional de Salud (National Institute of Health), Bogotá, Colombia.
Seventy clinical isolates of nontuberculous mycobacteria stored in 50% glycerol at -70 degrees C were identified by phenotypic techniques. The genotypic identification was made using the PCR-restriction analysis (PRA) using the restriction enzymes BstEII and HseIII, the restriction products were visualized on gels of agarose to 3%, and the concordance between the methodologies was evaluated.
A matching of 100% was obtained in the identification of Mycobacterium terrae, M. szulgai, M. avium, M. chelonae and M. scrofulaceum, the matching between M. fortuitum species, M. abscessus, M. gordonae and M. intracellulare varied from 44 to 89%; there was no concurrence in the identification of species M. flavescens and M. malmoense.
PRA provided a fast, inexpensive and accurate alternative for the identification of nontuberculous mycobacteria that permited the differentiation among species of a complex and determining the subtype of each species sample.
非结核分枝杆菌可以是腐生菌、致病菌或机会致病菌。这些微生物引起的最常见疾病包括麻醉手术后感染、与导管相关的感染、播散性皮肤病以及尤其影响艾滋病患者的肺部和中枢神经系统疾病。非结核分枝杆菌的鉴定可能需要数周时间，即便如此，也无法区分复杂菌群中的成员。
评估聚合酶链反应-限制性分析（PRA）技术作为一种对从哥伦比亚波哥大国家卫生研究所培养物保藏中心的临床样本中分离出的非结核分枝杆菌进行基因型鉴定的方法。
采用表型技术对保存在-70℃的50%甘油中的70株非结核分枝杆菌临床分离株进行鉴定。使用限制性内切酶BstEII和HseIII通过聚合酶链反应-限制性分析（PRA）进行基因型鉴定，将限制性产物在3%的琼脂糖凝胶上进行可视化，并评估两种方法之间的一致性。
在对土分枝杆菌、斯氏分枝杆菌、鸟分枝杆菌、龟分枝杆菌和瘰疬分枝杆菌的鉴定中获得了100%的匹配度，偶然分枝杆菌、脓肿分枝杆菌、戈登分枝杆菌和胞内分枝杆菌之间的匹配度在44%至89%之间；在微黄分枝杆菌和马尔默分枝杆菌的鉴定中没有一致性。
PRA为非结核分枝杆菌的鉴定提供了一种快速、廉价且准确的替代方法，该方法能够区分复杂菌群中的不同菌种并确定每个菌种样本的亚型。
