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. 1991 Sep;29(9):1985–1990. doi: 10.1128/jcm.29.9.1985-1990.1991

Detection of Pneumocystis carinii sequences by polymerase chain reaction: animal models and clinical application to noninvasive specimens.

K Kitada 1, S Oka 1, S Kimura 1, K Shimada 1, T Serikawa 1, J Yamada 1, H Tsunoo 1, K Egawa 1, Y Nakamura 1
PMCID: PMC270246  PMID: 1774325

Abstract

Pneumocystis carinii is a eukaryotic microbe which causes fatal pneumonia in patients with AIDS. Oligonucleotide primers were used to amplify the 5S rDNA sequence of P. carinii by the polymerase chain reaction (PCR) in various clinical and animal samples. Of 35 independent lung specimens tested, PCR detected the P. carinii sequence in all 23 cases which were known to be P. carinii infected, i.e., 15 from mice, 1 from rat, 3 from human autopsy, and 4 from biopsy of AIDS patients by needle aspiration. The results were consistent with clinical and microscopic diagnosis. The detection was highly sensitive and specific. Direct sequencing of these amplified DNAs revealed homogeneity of 5S rDNA sequences of independent isolates from mice, rats, and humans. Preliminary trials manifested efficacy of the PCR method to detect P. carinii sequences in induced sputum or blood from AIDS patients, the latter case suggesting that P. carinii might enter peripheral blood via phagocytosis or direct intrusion. Development of less-invasive or noninvasive PCR diagnostic techniques to detect P. carinii infection would greatly facilitate therapeutic and prophylactic management of P. carinii pneumonia.

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Selected References

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  1. Edman J. C., Kovacs J. A., Masur H., Santi D. V., Elwood H. J., Sogin M. L. Ribosomal RNA sequence shows Pneumocystis carinii to be a member of the fungi. Nature. 1988 Aug 11;334(6182):519–522. doi: 10.1038/334519a0. [DOI] [PubMed] [Google Scholar]
  2. Furuta T., Ueda K. Intra- and inter-species transmission and antigenic difference of Pneumocystis carinii derived from rat and mouse. Jpn J Exp Med. 1987 Feb;57(1):11–17. [PubMed] [Google Scholar]
  3. Gigliotti F., Stokes D. C., Cheatham A. B., Davis D. S., Hughes W. T. Development of murine monoclonal antibodies to Pneumocystis carinii. J Infect Dis. 1986 Aug;154(2):315–322. doi: 10.1093/infdis/154.2.315. [DOI] [PubMed] [Google Scholar]
  4. Goto H., Oka S., Mohri H., Kimura S., Mitamura K., Shimada K. Two cases of Pneumocystis carinii pneumonia occurred in human immunodeficiency virus infected patients: supplemental treatment with aerosolised pentamidine isethionate. Jpn J Med. 1989 Jan-Feb;28(1):105–109. doi: 10.2169/internalmedicine1962.28.105. [DOI] [PubMed] [Google Scholar]
  5. Mandal R. K. The organization and transcription of eukaryotic ribosomal RNA genes. Prog Nucleic Acid Res Mol Biol. 1984;31:115–160. doi: 10.1016/s0079-6603(08)60376-1. [DOI] [PubMed] [Google Scholar]
  6. Ng V. L., Gartner I., Weymouth L. A., Goodman C. D., Hopewell P. C., Hadley W. K. The use of mucolysed induced sputum for the identification of pulmonary pathogens associated with human immunodeficiency virus infection. Arch Pathol Lab Med. 1989 May;113(5):488–493. [PubMed] [Google Scholar]
  7. Pifer L. L., Woods D. R., Edwards C. C., Joyner R. E., Anderson F. J., Arheart K. Pneumocystis carinii serologic study in pediatric acquired immunodeficiency syndrome. Am J Dis Child. 1988 Jan;142(1):36–39. [PubMed] [Google Scholar]
  8. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  9. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Selik R. M., Starcher E. T., Curran J. W. Opportunistic diseases reported in AIDS patients: frequencies, associations, and trends. AIDS. 1987 Sep;1(3):175–182. [PubMed] [Google Scholar]
  11. Specht T., Wolters J., Erdmann V. A. Compilation of 5S rRNA and 5S rRNA gene sequences. Nucleic Acids Res. 1990 Apr 25;18 (Suppl):2215–2230. doi: 10.1093/nar/18.suppl.2215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Tanabe K., Fuchimoto M., Egawa K., Nakamura Y. Use of Pneumocystis carinii genomic DNA clones for DNA hybridization analysis of infected human lungs. J Infect Dis. 1988 Mar;157(3):593–596. doi: 10.1093/infdis/157.3.593. [DOI] [PubMed] [Google Scholar]
  13. Telzak E. E., Cote R. J., Gold J. W., Campbell S. W., Armstrong D. Extrapulmonary Pneumocystis carinii infections. Rev Infect Dis. 1990 May-Jun;12(3):380–386. doi: 10.1093/clinids/12.3.380. [DOI] [PubMed] [Google Scholar]
  14. Wakefield A. E., Pixley F. J., Banerji S., Sinclair K., Miller R. F., Moxon E. R., Hopkin J. M. Detection of Pneumocystis carinii with DNA amplification. Lancet. 1990 Aug 25;336(8713):451–453. doi: 10.1016/0140-6736(90)92008-6. [DOI] [PubMed] [Google Scholar]
  15. Walzer P. D. Diagnosis of Pneumocystis carinii pneumonia. J Infect Dis. 1988 Apr;157(4):629–632. doi: 10.1093/infdis/157.4.629. [DOI] [PubMed] [Google Scholar]
  16. Walzer P. D., Linke M. J. A comparison of the antigenic characteristics of rat and human Pneumocystis carinii by immunoblotting. J Immunol. 1987 Apr 1;138(7):2257–2265. [PubMed] [Google Scholar]
  17. Watanabe J., Hori H., Tanabe K., Nakamura Y. Phylogenetic association of Pneumocystis carinii with the 'Rhizopoda/Myxomycota/Zygomycota group' indicated by comparison of 5S ribosomal RNA sequences. Mol Biochem Parasitol. 1989 Jan 15;32(2-3):163–167. doi: 10.1016/0166-6851(89)90067-4. [DOI] [PubMed] [Google Scholar]
  18. Yoganathan T., Lin H., Buck G. A. An electrophoretic karyotype and assignment of ribosomal genes to resolved chromosomes of Pneumocystis carinii. Mol Microbiol. 1989 Nov;3(11):1473–1480. doi: 10.1111/j.1365-2958.1989.tb00132.x. [DOI] [PubMed] [Google Scholar]
  19. Yoshida Y. Ultrastructural studies of Pneumocystis carinii. J Protozool. 1989 Jan-Feb;36(1):53–60. doi: 10.1111/j.1550-7408.1989.tb02696.x. [DOI] [PubMed] [Google Scholar]

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