Rapid species identification of 391 clinicalMycobacterium isolates from Anhui Province by multi-locus PCR

The clinical Mycobacterium isolates were collected for the research to evaluate effect for rapid species identification.Primarily species were identified by culture method with differentiated PNB/TCH media.By means of multi-locus polymerase chain reaction(multi-locus PCR),the specific DNA fragments of 16SrRNA,Rv0577,IS1561,Rv1510,Rv1970,Rv3877/8 and Rv3120 genes in the genome of the isolates were examined respectively.The species were confirmed by rpoB-PRA and sequencing of hsp65 and rpoB.A total of 391 Mycobacterium isolates were collected for the species identification.By multi-locus PCR,378 were identified as Mycobacterium tuberculosis,6 were Mycobacterium africanum type I,and 7 were nontuberculous mycobacteria(NTM).The 7 NTM strains were confirmed to be one Mycobacterium avium,2 Mycobacterium massiliense,and 4 Mycobacterium intracellular.The results of multi-locus PCR were the same as that of rpoB-PRA and sequencing of hsp65 and rpoB.While by culture method with differentiated PNB/TCH media,385 strains were identified as Mycobacterium tuberculosis complex and 6 was NTM.Therefore,it's concluded that multi-locus PCR in this study could identify Mycobacterium species accurately,reliably,simply and rapidly.It has significantly useful value for the molecular epidemiology,clinical diagnosis and clinical therapy of Mycobacterium tuberculosis.