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Journal of Clinical Microbiology, September 2000, p. 3200-3204, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Molecular Characterization of Mycobacterium tuberculosis H37Rv/Ra Variants: Distinguishing the Mycobacterial Laboratory Strain†

P. Bifani,1,2 S. Moghazeh,1 B. Shopsin,1,2 J. Driscoll,3 A. Ravikovitch,1 and B. N. Kreiswirth1,*

Public Health Research Institute Tuberculosis Center1 and Department of Microbiology, New York University School of Medicine,2 New York, New York 10016, and Wadsworth Center, New York State Department of Health, Albany, New York 12201-20023

Received 22 December 1999/Returned for modification 23 February 2000/Accepted 16 May 2000

The Mycobacterium tuberculosis strains H37Rv and H37Ra are the most commonly used controls for M. tuberculosis identification in the clinical and research laboratory setting. To reduce the likelihood of misidentification and possible cross-contamination with this laboratory neotype, it is important to be able to distinguish H37 from clinical isolates. To provide a reference for identifying H37, we used multiple molecular techniques to characterize H37 strains, including 18 of the most frequently used variants available through the American Type Culture Collection. Isolates were genotyped using gene probes to IS6110 and IS1085. In addition, we performed polymorphic GC-rich sequence typing (PGRS), spoligotyping, determination of variable number of tandem repeats (VNTR), and PCR amplification of the mtp40, msx4, and mpp8 polymorphic regions. Southern hybridization with IS6110 provided the most discrimination, differentiating the 18 H37 isolates into 10 discrete patterns made up of 9 H37Rv variants and 1 H37Ra variant. PGRS, IS1085, mpp8, and spoligotyping were not able to distinguish any H37 variants, while VNTR and msx4 discriminated two. Only IS6110 and spoligotyping could distinguish the H37 strain from clinical isolates. In summary, spoligotyping and IS6110 provide a rapid and accurate way to identify H37 contamination, though IS6110 can, in addition, classify many of the H37 variants that would otherwise require phenotypic segregation.

* Corresponding author. Mailing address: Public Health Research Institute Tuberculosis Center, 455 First Ave., New York, NY 10016. Phone: (212) 578-0850. Fax: (212) 578-0853. E-mail: barry{at}phri.nyu.edu.

dagger Publication 72 from the Public Health Research Institute Tuberculosis Center.

Journal of Clinical Microbiology, September 2000, p. 3200-3204, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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