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Journal of Clinical Microbiology, July 2003, p. 2822-2826, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.2822-2826.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Use of Molecular Methods To Identify the Mycobacterium tuberculosis Complex (MTBC) and Other Mycobacterial Species and To Detect Rifampin Resistance in MTBC Isolates following Growth Detection with the BACTEC MGIT 960 System

Akos Somoskovi,1,2 Qunfeng Song,1,3 Judit Mester,1,4 Charise Tanner,5 Yvonne M. Hale,5 Linda M. Parsons,1,6 and Max Salfinger1,6,7*

Wadsworth Center, New York State Department of Health,1 Department of Biomedical Sciences, School of Public Health, University at Albany,6 Department of Medicine, Albany Medical College, Albany, New York,7 Department of Respiratory Medicine, School of Medicine, Semmelweis University,2 Korányi National Institute for Tuberculosis and Respiratory Medicine, Budapest, Hungary,4 Guizhou Provincial Epidemic Preventive Station, Guiyang, China,3 Bureau of Laboratories, Florida Department of Health, Jacksonville, Florida5

Received 29 January 2003/ Returned for modification 10 March 2003/ Accepted 14 April 2003

A prospective study was organized by using a total of 1,585 consecutive clinical specimens to determine whether biomass obtained from positive growth in the MGIT 960 system could be used directly in AccuProbe DNA hybridization tests, the PCR-based Inno-LiPA Rif.TB (LiPA) assay, and a PCR-based DNA sequencing of the rpoB gene for the rapid identification of the Mycobacterium tuberculosis complex (MTBC) and other mycobacterial species and for the determination of rifampin (RIF) resistance in MTBC strains. The results were compared to routine culture, identification, and susceptibility testing techniques performed on the same samples. The study results revealed that the DNA AccuProbe assay (on the day of growth positivity) readily identified 95.7%, the LiPA assay readily identified 98.6%, and rpoB sequencing readily identified 97.1% of the 70 MTBC isolates from mycobacterial growth indicator tubes (MGIT). In addition, application of the LiPA for the identification and RIF susceptibility testing of the MTBC in growth-positive MGIT resulted in a turnaround time of less than 2 weeks after specimen receipt. Although DNA sequencing of rpoB required a slightly longer (16 days) turnaround time, this method was capable of identifying several species of nontuberculous mycobacteria in addition to identifying MTBC and determining RIF susceptibility or resistance. The molecular methods were also found to rapidly identify RIF-susceptible and -resistant MTBC in two of the three mixed mycobacterial cultures weeks earlier than conventional methods. In conclusion, the biomass obtained in MGIT at the time of growth positivity in the 960 system is sufficient for use in all three molecular tests, and this approach can reduce the turnaround time for reporting results.

* Corresponding author. Mailing address: Wadsworth Center, New York State Department of Health, P.O. Box 509, Albany, NY 12201-0509. Phone: (518) 474-2196. Fax: (518) 474-6964. E-mail: salfinger{at}wadsworth.org.

Journal of Clinical Microbiology, July 2003, p. 2822-2826, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.2822-2826.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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