Victor T C, Jordaan A M, Van Schalkwyk E J, Coetzee G J, Van Helden P D
MRC Centre for Molecular and Cellular Biology, Department of Medical Physiology and Biochemistry, University of Stellenbosch Medical School, South Africa.
J Med Microbiol. 1996 May;44(5):332-9. doi: 10.1099/00222615-44-5-332.
The polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique was evaluated for species identification among mycobacteria by analysis of the dnaJ gene. Nine clinical isolates of Mycobacterium tuberculosis with different fingerprint patterns all gave the same distinct SSCP banding pattern and could be distinguished from other mycobacteria, such as M. avium. In contrast, considerable strain-specific dnaJ gene variations were observed amongst 42 clinical isolates of M. avium and 13 other atypical mycobacterial strains. Only 62% of the M. avium isolates hybridised to an M. avium-specific probe and only 14% could be identified correctly as M. avium by both probe and restriction fragment length polymorphism analysis. This finding was supported by direct sequence analysis. Variations were also observed in M. gordonae and M. scrofulaceum isolates. Computerised analysis of M. avium samples broadly identified three clusters. Results suggest that although the SSCP procedure may be useful for distinguishing M. tuberculosis from other mycobacteria, this technique applied to the dnaJ gene may not be suitable for strain identification. The results stress the importance of testing a large collection of clinical isolates before new molecular procedures are introduced into routine laboratories.
通过对dnaJ基因的分析,评估聚合酶链反应-单链构象多态性(PCR-SSCP)技术在分枝杆菌菌种鉴定中的应用。9株具有不同指纹图谱的结核分枝杆菌临床分离株均呈现相同的独特SSCP条带模式,且可与其他分枝杆菌如鸟分枝杆菌区分开来。相比之下,在42株鸟分枝杆菌临床分离株和13株其他非典型分枝杆菌菌株中观察到了相当多的菌株特异性dnaJ基因变异。仅62%的鸟分枝杆菌分离株与鸟分枝杆菌特异性探针杂交,通过探针和限制性片段长度多态性分析,只有14%能够被正确鉴定为鸟分枝杆菌。直接测序分析支持了这一发现。在戈登分枝杆菌和瘰疬分枝杆菌分离株中也观察到了变异。对鸟分枝杆菌样本的计算机分析大致确定了三个聚类。结果表明,虽然SSCP程序可能有助于区分结核分枝杆菌与其他分枝杆菌,但应用于dnaJ基因的这项技术可能不适用于菌株鉴定。结果强调了在将新的分子程序引入常规实验室之前,对大量临床分离株进行检测的重要性。