Detection and identification of Mycobacterium directly from Bactec bottles by using a DNA-rRNA probe

Abstract

Detection and identification of mycobacterial species using DNA-rRNA probes from colonies has been used for some time. Guidelines for probe use with a specific minimum growth index (GI) cutoff from Bactec 12B bottles or its use with 7H9 broth has not been defined. This study defined this minimum GI guideline. Clinical specimens received during the study period were decontaminated and directly inoculated into Bactec 12B bottles and appropriate solid media. An initial probe test was performed at a GI reading of ⩾100. An additional probe test was made at GI of ⩾300. A total of 1377 specimens were processed with 98 specimens containing 99 isolates of Mycobacterium. Of these, 82 were M. avium complex (MAC), six were M. tuberculosis (Mtb), and 11 were other species. At a GI of ⩾100, 82% of MAC were detected; at a GI of ⩾300, 89% were detected. All Mtb were detected at a GI of ⩾100. The probes correctly identified all isolates from backup 7H9 broths. There were no false-positive probe results for any of the isolates from either Bactec 12B or 7H9 media. Therefore, the DNA-rRNA probe test at a GI of ⩾300 saves considerable time (on average, 21 days saved) for the detection and identification of Mtb and MAC. This time saving compares with waiting for the production of colonies while retaining acceptable sensitivity and excellent specificity.