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

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, January 2007, p. 173-178, Vol. 45, No. 1
0095-1137/07/$08.00+0     doi:10.1128/JCM.01776-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Detection of Coccidioides Species in Clinical Specimens by Real-Time PCR{triangledown}

M. J. Binnicker,1 S. P. Buckwalter,1 J. J. Eisberner,1 R. A. Stewart,2 A. E. McCullough,2 S. L. Wohlfiel,1 and N. L. Wengenack1*

Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic and Mayo Clinic College of Medicine, Rochester, Minnesota 55905,1 Department of Laboratory Medicine and Pathology, Mayo Clinic Arizona, Scottsdale, Arizona 852592

Received 28 August 2006/ Returned for modification 13 September 2006/ Accepted 16 October 2006

Coccidioides spp. are dimorphic fungal pathogens endemic to the semiarid regions of North, Central, and South America. Currently, direct smear and culture are the most common means of identifying Coccidioides spp. While these methods offer relatively sensitive and specific means of detecting Coccidioides spp., growth in culture may take up to 3 weeks, potentially delaying the diagnosis and initiation of appropriate antifungal therapy. In addition, growth of the organism represents a significant safety risk to laboratory personnel. The need for a rapid and safe means of diagnosing coccidioidomycosis prompted us to develop a real-time PCR assay to detect Coccidioides spp. directly from clinical specimens. Primers and fluorescent resonance energy transfer (FRET) probes were designed to target the internal transcribed spacer 2 region of Coccidioides. The assay's limit of detection is below 50 targets per reaction. An analysis of 40 Coccidioides sp. clinical isolates grown in culture demonstrated 100% sensitivity of the assay. A cross-reactivity panel containing fungi, bacteria, mycobacteria, and viruses was tested and demonstrated 100% specificity for Coccidioides spp. An analysis of 266 respiratory specimens by LightCycler PCR demonstrated 100% sensitivity and 98.4% specificity for Coccidioides spp. compared with culture. Analysis of 66 fresh tissue specimens yielded 92.9% sensitivity and 98.1% specificity versus those of the culture method. The sensitivity of the assay testing 148 paraffin-embedded tissue samples is 73.4%. A rapid method for the detection of Coccidioides spp. directly from clinical material will greatly assist in the timely diagnosis and treatment of patients, while at the same time decreasing the risk of accidental exposure to laboratory personnel.

* Corresponding author. Mailing address: Mayo Clinic, 200 First Street SW Hilton 860A, Rochester, MN 55905. Phone: (507) 266-4822. Fax: (507) 284-4272. E-mail: wengenack.nancy{at}mayo.edu.

{triangledown} Published ahead of print on 15 November 2006.

Journal of Clinical Microbiology, January 2007, p. 173-178, Vol. 45, No. 1
0095-1137/07/$08.00+0     doi:10.1128/JCM.01776-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hombach, M., Stulz, P., Arnold, W., Pfyffer, G. E. (2008). Pulmonary Coccidioides nodule in a Swiss patient with chronic lymphatic leukaemia. J Med Microbiol 57: 1427-1430 [Abstract] [Full Text]  
  • Vollmer, T., Stormer, M., Kleesiek, K., Dreier, J. (2008). Evaluation of Novel Broad-Range Real-Time PCR Assay for Rapid Detection of Human Pathogenic Fungi in Various Clinical Specimens. J. Clin. Microbiol. 46: 1919-1926 [Abstract] [Full Text]  
  • Parish, J. M., Blair, J. E. (2008). Coccidioidomycosis. Mayo Clin Proc. 83: 343-349 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»