This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Cloud, J. L. Articles by Woods, G. L. Search for Related Content PubMed PubMed Citation Articles by Cloud, J. L. Articles by Woods, G. L.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, August 2005, p. 4205-4207, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.4205-4207.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Use of the MGB Eclipse System and SmartCycler PCR for Differentiation of Mycobacterium chelonae and M. abscessus

Joann L. Cloud,^1* Karen Hoggan,¹ Evgeniy Belousov,² Samuel Cohen,³ Barbara A. Brown-Elliott,⁴ Linda Mann,⁴ Rebecca Wilson,⁴ Wade Aldous,^1,3,5 Richard J. Wallace Jr.,⁴ and Gail L. Woods^1,3,5

ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, Utah,¹ Nanogen, Inc., Bothell, Washington,² ARUP Laboratories, Salt Lake City, Utah,³ University of Texas Health Center, Tyler, Texas,⁴ University of Utah, Department of Pathology, Salt Lake City, Utah⁵

Received 18 March 2005/ Returned for modification 10 May 2005/ Accepted 17 May 2005

Although accurate in the identification of Mycobacterium species, partial 16S rRNA gene sequencing does not distinguish Mycobacterium chelonae from M. abscessus. Thus, we designed a SmartCycler PCR assay targeting the 16S-to-23S internal transcribed spacer (ITS) region with use of MGB Eclipse probes to distinguish each species. Comparison with PCR-restriction enzyme analysis of a 441-bp fragment of the hsp65 gene resulted in 100% correlation with 25 isolates of M. chelonae and 25 isolates of M. abscessus. ITS PCR performed on 90 consecutive isolates identified by partial 16S rRNA gene sequencing (26 isolates of the M. chelonae-M. abscessus complex and 64 remaining isolates, including Mycobacterium species, Nocardia species, and other aerobic actinomycetes) showed 100% specificity and sensitivity. The ITS PCR assay is accurate and specific, easy to perform, and a good supplemental test when using partial 16S rRNA gene sequencing to identify M. chelonae and M. abscessus.

---

* Corresponding author. Mailing address: ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT 84108. Phone: (801) 583-2787, ext. 2439. Fax: (801) 584-5207. E-mail: cloudjl{at}aruplab.com.

---

Journal of Clinical Microbiology, August 2005, p. 4205-4207, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.4205-4207.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Woo, P. C. Y., Teng, J. L. L., Wu, J. K. L., Leung, F. P. S., Tse, H., Fung, A. M. Y., Lau, S. K. P., Yuen, K.-y. (2009). Guidelines for interpretation of 16S rRNA gene sequence-based results for identification of medically important aerobic Gram-positive bacteria. J Med Microbiol 58: 1030-1036 [Abstract] [Full Text]  
• Simmon, K. E., Pounder, J. I., Greene, J. N., Walsh, F., Anderson, C. M., Cohen, S., Petti, C. A. (2007). Identification of an Emerging Pathogen, Mycobacterium massiliense, by rpoB Sequencing of Clinical Isolates Collected in the United States. J. Clin. Microbiol. 45: 1978-1980 [Abstract] [Full Text]  
• Neal, H., Cloud, J. L., Pounder, J. I., Page, S. R., Woods, G. L. (2005). Sequence Variant for Internal Transcribed Spacer Region of Mycobacterium abscessus. J. Clin. Microbiol. 43: 6214-6214 [Full Text]