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Journal of Clinical Microbiology, May 2002, p. 1739-1742, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1739-1742.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Rapid Identification of Penicillium marneffei by PCR-Based Detection of Specific Sequences on the rRNA Gene

Nongnuch Vanittanakom,1* Pramote Vanittanakom,2 and Roderick J. Hay3

Department of Microbiology,1 Department of Pathology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand,2 St. John's Institute of Dermatology, Guy's, King's and St. Thomas’ School of Medicine, University of London, United Kingdom3

Received 19 June 2001/ Returned for modification 12 July 2001/ Accepted 15 February 2002

An emerging pathogenic dimorphic fungus, Penicillium marneffei, is one of the major causes of morbidity in patients with human immunodeficiency virus infection in Southeast Asia. A PCR-hybridization assay has been developed to identify this pathogen. This study describes the use of single and nested PCR methods for the rapid identification of P. marneffei. Two sets of oligonucleotide primers were derived from the sequence of 18S rRNA genes of P. marneffei. The outer primers (RRF1 and RRH1) were fungus specific. The inner primers (Pm1 and Pm2) were specific for P. marneffei and were used in nested or single PCR. The specific fragment of approximately 400-bp was amplified from both mold and yeast forms of 13 P. marneffei human isolates, 12 bamboo rat isolates, and 1 soil isolate, but not from other fungi, bacteria, and human DNA. The amplified products were analyzed by agarose gel electrophoresis followed by ethidium bromide staining. The sensitivities of the single PCR and nested PCR were 1.0 pg/µl and 1.8 fg/µl, respectively. The assay is useful for rapid identification of P. marneffei cultures. Very young culture of P. marneffei (2-day-old filamentous colony, 2 mm in diameter) could be performed by this assay. The species was identified within 7 h (single PCR) or 10 h (nested PCR), compared to 4 to 7 days for confirmation of dimorphism. The application of these PCR methods for early diagnosis of the disease needs to be studied further.

* Corresponding author. Mailing address: Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand. Phone: (66) 53 945332, ext. 412. Fax: (66) 53 217144. E-mail: nvanitta{at}mail.med.cmu.ac.th.

Journal of Clinical Microbiology, May 2002, p. 1739-1742, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1739-1742.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Thirach, S., Cooper, C. R. Jr, Vanittanakom, N. (2008). Molecular analysis of the Penicillium marneffei glyceraldehyde-3-phosphate dehydrogenase-encoding gene (gpdA) and differential expression of gpdA and the isocitrate lyase-encoding gene (acuD) upon internalization by murine macrophages. J Med Microbiol 57: 1322-1328 [Abstract] [Full Text]  
  • Vanittanakom, N., Cooper, C. R. Jr., Fisher, M. C., Sirisanthana, T. (2006). Penicillium marneffei Infection and Recent Advances in the Epidemiology and Molecular Biology Aspects. Clin. Microbiol. Rev. 19: 95-110 [Abstract] [Full Text]  
  • Guedes, H. L. d. M., Guimaraes, A. J., Muniz, M. d. M., Pizzini, C. V., Hamilton, A. J., Peralta, J. M., Deepe, G. S. Jr., Zancope-Oliveira, R. M. (2003). PCR Assay for Identification of Histoplasma capsulatum Based on the Nucleotide Sequence of the M Antigen. J. Clin. Microbiol. 41: 535-539 [Abstract] [Full Text]  


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