評估Multiplex PCR在Mycobacterium Tuberculosis Complex快速鑑定的應用

Objective: Tuberculosis has been one of the major causes of death in Taiwan for decades. Infection with Non-tuberculous mycobacteria（NTM）are increasingly observed in immunocomprosied patients in these years, however. Mycobacterium tuberculosis complex, the causative agents of human TB is the commonest pathogen for pulmonary and extra pulmonary tuberculosis cases. Although there are many techniques available for rapid detection of DNA of members of the MTBC, culture is still gold standard to confirm the presence of 'live' microorganisms from suspected patients' specimens. The strain identification of Mycobacterium was based on culture and conventional phenotypic methods which are cumbersome, labor intensive procedures and time-consuming. We have developed a multiplex PCR in these years and implemented the rapid strain identification in a single tube. The result of the study show that the multiplex PCR is supposed to be less time-consuming, less complicated than conventional methods. We developed a multiplex PCR based on amplification of 365, 541 and 165 bp fragments of dnaJ, IS611 and hsp65. The multiplex PCR was tested with 379 clinical isolates. The results of the study show that 141 isolates of M. tuberculosis 238 isolates of NTM are confirmed. The sensitivity and the specificity of the multiplex PCR is 100%and 98.7% by comparison with PCR-RFLP. These results suggest that multiplex PCR multiplex PCR could be used as less complicated, less time consuming and cost-effective method for the rapid identification of MTBC.