Cormican M, Glennon M, Riain U N, Flynn J
Department of Medical Microbiology, University College Hospital, Galway, Ireland.
J Clin Pathol. 1995 Mar;48(3):203-5. doi: 10.1136/jcp.48.3.203.
To develop a multiplex polymerase chain reaction (PCR) method to facilitate identification of mycobacterial isolates.
Type strains of 14 species of mycobacteria and 56 clinical isolates were lysed by boiling in TE Triton. The lysate (5 microliters) was used directly in a PCR reaction incorporating three pairs of PCR primers expected to amplify fragments from the genome of (a) all mycobacteria, (b) Mycobacterium tuberculosis complex only and (c) M avium only. PCR products were visualised by electrophoresis on agarose gels.
Multiplex PCR applied to 14 type strains yielded patterns on electrophoresis which permitted identification of the mycobacterial isolates as M tuberculosis complex, M avium or as mycobacteria other than the former. The identification of 56 clinical isolates by multiplex PCR was consistent with other methods and was accomplished in less than one working day.
This method may facilitate rapid and convenient identification of most clinical isolates of mycobacteria by PCR and gel electrophoresis. Further evaluation is warranted.
开发一种多重聚合酶链反应（PCR）方法，以促进分枝杆菌分离株的鉴定。
将14种分枝杆菌的标准菌株和56株临床分离株在含有吐温的TE缓冲液中煮沸裂解。取5微升裂解物直接用于PCR反应，该反应包含三对PCR引物，预期可从以下基因组中扩增片段：（a）所有分枝杆菌，（b）仅结核分枝杆菌复合群，（c）仅鸟分枝杆菌。PCR产物通过琼脂糖凝胶电泳进行可视化分析。
对14种标准菌株进行多重PCR后，电泳图谱可将分枝杆菌分离株鉴定为结核分枝杆菌复合群、鸟分枝杆菌或其他分枝杆菌。通过多重PCR对56株临床分离株的鉴定结果与其他方法一致，且在不到一个工作日内即可完成。
该方法可通过PCR和凝胶电泳快速便捷地鉴定大多数分枝杆菌临床分离株。值得进一步评估。
