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 McNabb, A. Articles by Isaac-Renton, J. Search for Related Content PubMed PubMed Citation Articles by McNabb, A. Articles by Isaac-Renton, J.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, January 2006, p. 60-66, Vol. 44, No. 1
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.1.60-66.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Direct Identification of Mycobacteria in Primary Liquid Detection Media by Partial Sequencing of the 65-Kilodalton Heat Shock Protein Gene

Alan McNabb,^1* Kathy Adie,¹ Mabel Rodrigues,¹ William A. Black,^1,2 and Judith Isaac-Renton^1,2

Laboratory Services, British Columbia Centre for Disease Control,¹ Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada²

Received 22 April 2005/ Returned for modification 1 August 2005/ Accepted 23 October 2005

We investigated extending the use of direct partial hsp65 gene sequencing for the identification of mycobacteria to isolates in primary liquid detection media as an economical, feasible, and more rapid means of identification. During the course of the study, the hsp65 sequence-based identifications for isolates from 670 primary liquid detection media determined to be positive for acid-fast bacilli were compared to the identifications derived from Accuprobes, biochemical test panels, or 16S rRNA gene sequencing. Preliminary analysis indicated a 97.6% concordance, with a final agreement of 99.1% between the identification algorithms. hsp65 sequencing costs (US$32.84) were greater than the cost of identification with Accuprobe (US$19) but less than the cost of the biochemical test panel identification (average cost, US$98.90) and equivalent to the cost of 16S rRNA sequencing, although there was a referral cost (US$59.85) for the shipping of isolates to another reference laboratory. Analysis indicated that our laboratory would have recognized a cost savings of approximately $12,000 by using hsp65 sequencing to identify isolates from specimens with a negative fluorescent- smear status and would have achieved further savings by using it as an alternative to biochemical panel testing for fluorescent-smear-positive specimens. The time to identification by hsp65 gene sequencing was slightly longer than that required by the Accuprobe assay (1 versus 2 days), shorter than that required by the biochemical test panels (2 days versus 26 days on average), and more rapid than referral for 16S rRNA gene sequencing.

---

* Corresponding author. Mailing address: Molecular Services, British Columbia Centre for Disease Control, 655 West 12th Avenue, Vancouver, British Columbia V5Z 4R4, Canada. Phone: (604) 660-0876. Fax (604) 660-6073. E-mail alan.mcnabbc{at}bccdc.ca.

---

Journal of Clinical Microbiology, January 2006, p. 60-66, Vol. 44, No. 1
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.1.60-66.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Leader, B. T., Frye, J. G., Hu, J., Fedorka-Cray, P. J., Boyle, D. S. (2009). High-Throughput Molecular Determination of Salmonella enterica Serovars by Use of Multiplex PCR and Capillary Electrophoresis Analysis. J. Clin. Microbiol. 47: 1290-1299 [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]