Application of multiplex PCR in rapid identification of Mycobacterium tuberculosis and non-tuberculosis Mycobacteria / 解放军医学杂志

ABSTRACT

Objective To investigate the feasibility of multiplex PCR for rapid identification of Mycobacterium tuberculosis and non-tuberculosis Mycobacteria. Methods According to MTP40 gene sequence of Mycobacterium tuberculosis, 32kD gene sequence of Mycobacterium and IS6110 insertion sequence gene sequence of Mycobacterium tuberculosis complex, three specific pairs of primers (PT1-PT2, MT1-MT2 and IS5-IS6) for Mycobacterium were designed, and the target DNA for MTP40, 32kD and IS6110 was 396bp, 506bp and 984bp, respectively. The genome of 92 Mycobacterium tuberculosis clinically isolated strains and 5 non-tuberculosis Mycobacteria clinical strains were amplified in the same system, and the results were compared with reference strains. Results Among 92 clinical strains of Mycobacterium tuberculosis, the DNA fragments of 396bp, 506bp and 984bp were found in 90 Mycobacterium tuberculosis clinical strains, as well as in the reference strain H37Rv; the sensitivity of multiplex PCR for Mycobacterium tuberculosis was 97.8%, and the specificity was 100.0%. The DNA fragments of 506bp were all found in 5 non-tuberculosis Mycobacteria clinical strains, the sensitivity and specificity for non-tuberculosis Mycobacterium were both 100.0%. Conclusion The multiplex PCR is a rapid, sensitive and specific method for identification of Mycobacterium tuberculosis and non-tuberculosis Mycobacteria, and it may provide an effective way for clinical diagnosis of Mycobacterium tuberculosis and non-tuberculosis Mycobacteria, therefore useful in clinical application.