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Journal of Clinical Microbiology, October 2001, p. 3437-3441, Vol. 39, No. 10
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.10.3437-3441.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Widespread Occurrence of Pneumocystis carinii in Commercial Rat Colonies Detected Using Targeted PCR and Oral Swabs

Crystal R. Icenhour, Sandra L. Rebholz, Margaret S. Collins, and Melanie T. Cushion*

Department of Infectious Disease, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267-0560, and VA Medical Center, Research 151, Cincinnati, Ohio 45220

Received 18 January 2001/Returned for modification 27 March 2001/Accepted 27 June 2001

The genus Pneumocystis contains a family of fungal organisms that infect a wide variety of mammalian species. Although it is a cause of pneumonia in immunocompromised hosts, recent evidence suggests that these organisms colonize nonimmunosuppressed hosts. Detection of cryptic colonization with Pneumocystis becomes important in animal studies when infection-free animals are necessary. Provocation by chronic immunosuppression, histology, and serology has been widely used to detect the presence of Pneumocystis in rat colonies, requiring lengthy time periods and/or postmortem tissue. We conducted a study to evaluate the use of PCR amplification of oral swabs for the antemortem detection of Pneumocystis in 12 rat groups from three commercial vendors. Sera were collected upon arrival, and the oral cavity was swabbed for PCR analysis. Ten of these groups of rats were then housed in pairs under barrier and immunosuppressed to provoke Pneumocystis growth. Once moribund, the rats were sacrificed, and the lungs were collected to evaluate the presence of Pneumocystis by PCR and microscopic enumeration. DNA was extracted from oral swabs and lung homogenates, and PCR was performed using primers targeting a region within the mitochondrial large-subunit rRNA of Pneumocystis carinii f. sp. carinii. Upon receipt, 64% of rats were positive for P. carinii f. sp. carinii-specific antibodies, while P. carinii f. sp. carinii DNA was amplified from 98% of oral swabs. Postmortem PCR analysis of individual lungs revealed P. carinii f. sp. carinii DNA in all rat lungs, illustrating widespread occurrence of Pneumocystis in commercial rat colonies. Thus, oral swab/PCR is a rapid, nonlethal, and sensitive method for the assessment of Pneumocystis exposure.

* Corresponding author. Mailing address: Dept. of Infectious Disease, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0560. Phone: (513) 861-3100, ext. 4417. Fax: (513) 475-6415. E-mail: Melanie.Cushion{at}uc.edu.

Journal of Clinical Microbiology, October 2001, p. 3437-3441, Vol. 39, No. 10
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.10.3437-3441.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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