Development and evaluation of the internal-controlled real-time PCR assay for Rhodococcus equi detection in various clinical specimens
We investigated the use of multiprimer-PCR for detection of mycobacteria species in clinical samples. Three different mycobacterial genomic fragments were investigated: the IS6110 insertion sequence, present in M. tuberculosis complex; the genus specific fragment (32kDa); and from M. tuberculosis species-specific mtp40 gene. The sensitivity and specificity using 135 clinical isolates were 94.5% and 95.9%, respectively, compared with culture in Löwenstein-Jensen medium; the detection limit was 0.05ng of DNA. In conclusion, this assay is reliable and rapid for detection of Mycobacterium species in clinical samples, and differentiates M. tuberculosis from M. bovis strains in a single-step assay.