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Journal of Clinical Microbiology, September 1998, p. 2491-2494, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Determination of Copy Number of rRNA Genes in Pneumocystis carinii f. sp. hominis

Xing Tang,1 Marilyn S. Bartlett,1 James W. Smith,1 Jang-Jih Lu,2 and Chao-Hung Lee1,*

Departments of Pathology and Laboratory Medicine, Indiana University School of Medicine, Indianapolis, Indiana 46202,1 and Department of Pathology, Tri-Service General Hospital and National Defense Medical Center, Taipei, Taiwan2

Received 2 March 1998/Returned for modification 29 April 1998/Accepted 13 June 1998

Differential PCR was performed to determine the copy number of rRNA genes in Pneumocystis carinii f. sp. hominis. Two different reference genes, thymidylate synthase (TS) and beta-tubulin (BTU) genes, were used. Primers for the internal transcribed spacer (ITS) region of nuclear rRNA genes and either the TS or BTU gene were mixed together to perform PCR on seven different bronchoalveolar lavage specimens from patients with P. carinii pneumonia. The radioactivity derived from the incorporated radioactive nucleotides of each PCR product band was then used to calculate the copy number of the ITS relative to that of the TS or BTU gene. The copy number ratio between the ITS and the TS gene was determined to be 0.8, and that between the ITS and the BTU gene was also 0.8. These results suggest that the ITS has the same copy number as the TS or BTU gene. Since the copy number of the TS or BTU gene is presumed to be 1, the results also suggest that P. carinii f. sp. hominis has only one copy of the ITS and thus one copy of the nuclear rRNA genes. Therefore, two types of ITS sequences derived from a specimen would indicate that the patient is infected by two types of P. carinii f. sp. hominis.

* Corresponding author. Mailing address: Department of Pathology and Laboratory Medicine, Indiana University School of Medicine, 1120 South Dr., FH 419, Indianapolis, IN 46202-5113. Phone: (317) 274-2596. Fax: (317) 278-0643. E-mail: chlee{at}iupui.edu.

Journal of Clinical Microbiology, September 1998, p. 2491-2494, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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