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Journal of Clinical Microbiology, July 1998, p. 2068-2072, Vol. 36, No. 7
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Diagnostic Use of PCR for Detection of Pneumocystis carinii in Oral Wash Samples

Jannik Helweg-Larsen,1,3 Jørgen Skov Jensen,2 Thomas Benfield,3 Ulrik Gerner Svendsen,4 Jens D. Lundgren,3 and Bettina Lundgren5,*

Departments of Clinical Microbiology1 and Infectious Diseases,3 Hvidovre Hospital, Neisseria Department2 and Department of Clinical Microbiology,5 Statens Serum Institut, and Department of Cardiology, Rigshospitalet,4 Copenhagen, Denmark

Received 30 December 1997/Returned for modification 12 February 1998/Accepted 3 April 1998

There is a need to develop noninvasive methods for the diagnosis of Pneumocystis carinii pneumonia in patients unable to undergo bronchoscopy or induction sputum. Oral wash specimens are easily obtained, and P. ca- rinii nucleic acid can be amplified and demonstrated by PCR. In routine clinical use, easy sample processing and single-round PCR are needed to ensure rapid analysis and to reduce the risk of contamination. We developed a single-round Touchdown PCR (TD-PCR) protocol with the ability to detect PCR inhibition in the specimen. The TD-PCR was evaluated in a routine diagnostic laboratory and was compared to a previously described PCR protocol (mitochondrial RNA) run in a research laboratory. Both PCR methods amplified a sequence of the mitochondrial rRNA gene of P. carinii. Paired bronchoalveolar lavage (BAL) and oral wash specimens from 76 consecutive human immunodeficiency virus type 1-infected persons undergoing a diagnostic bronchoscopy were included. The TD-PCR procedure was quicker than the mitochondrial PCR procedure (<24 versus 48 h) and, compared to microscopy, had sensitivity, specificity, and positive and negative predictive values of 89, 94, 93, and 91%, respectively, for oral wash specimens and 100, 91, 90, and 100%, respectively, for BAL specimens. Our results suggest that oral wash specimens are a potential noninvasive method to obtain a diagnostic specimen during P. carinii pneumonia infection and that it can be applied in a routine diagnostic laboratory.

* Corresponding author. Present address: Department of Infectious Diseases, Hvidovre Hospital, 2650 Hvidovre, Denmark. Phone: 45 36 32 30 15. Fax: 45 36 47 33 40. E-mail: eurosida{at}inet.uni2.dk.

Journal of Clinical Microbiology, July 1998, p. 2068-2072, Vol. 36, No. 7
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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