Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Mar;33(3):718–724. doi: 10.1128/jcm.33.3.718-724.1995

Use of semiquantitative PCR to assess onset and treatment of Pneumocystis carinii infection in rat model.

T J O'Leary 1, M M Tsai 1, C F Wright 1, M T Cushion 1
PMCID: PMC228020  PMID: 7751383

Abstract

The use of semiquantitative PCR (SQPCR) to assess Pneumocystis carinii pneumonia (PCP) infection and its response to treatment was studied with rats. Groups of eight rats were immunosuppressed with steroids for 3 to 12 weeks. Untreated controls were maintained for the same periods. Three groups of rats were treated with pentamidine, three groups were treated with trimethoprim-sulfamethoxazole, and three groups of rats were tapered from steroids. At various times during suppression, rats from the different groups were sacrificed. At necropsy, lungs were lavaged to obtain bronchoalveolar fluids and then homogenized. Bronchoalveolar fluids and homogenates were assayed by cyst counting and SQPCR. An increase in the SQPCR signal was seen throughout immunosuppression, with a slow decrease upon the withdrawal of steroids and a faster decrease with drug treatment. SQPCR results with lung homogenates and bronchoalveolar fluids strongly correlated with each other and with cyst counts. These results warrant investigation of SQPCR for assessing treatment results of human P. carinii pneumonia infection.

Full Text

The Full Text of this article is available as a PDF (247.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amin M. B., Mezger E., Zarbo R. J. Detection of Pneumocystis carinii. Comparative study of monoclonal antibody and silver staining. Am J Clin Pathol. 1992 Jul;98(1):13–18. doi: 10.1093/ajcp/98.1.13. [DOI] [PubMed] [Google Scholar]
  2. Armstrong D., Bernard E. M. Human immunodeficiency virus (HIV)-associated pneumocystosis. Curr Clin Top Infect Dis. 1991;11:19–34. [PubMed] [Google Scholar]
  3. Bartlett M. S., Smith J. W. Pneumocystis carinii, an opportunist in immunocompromised patients. Clin Microbiol Rev. 1991 Apr;4(2):137–149. doi: 10.1128/cmr.4.2.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blumenfeld W., McCook O., Holodniy M., Katzenstein D. A. Correlation of morphologic diagnosis of Pneumocystis carinii with the presence of pneumocystis DNA amplified by the polymerase chain reaction. Mod Pathol. 1992 Mar;5(2):103–106. [PubMed] [Google Scholar]
  5. Blumenfeld W., Miller C. N., Chew K. L., Mayall B. H., Griffiss J. M. Correlation of Pneumocystis carinii cyst density with mortality in patients with acquired immunodeficiency syndrome and pneumocystis pneumonia. Hum Pathol. 1992 Jun;23(6):612–618. doi: 10.1016/0046-8177(92)90315-t. [DOI] [PubMed] [Google Scholar]
  6. Borensztein L., Hatin I., Simonpoli A. M., Ugarte E., Girard P. M., Jaureguiberry G. An alternative to DNA extraction for the diagnosis of Pneumocystis carinii pneumonia by polymerase chain reaction using a new oligonucleotide probe. Mol Cell Probes. 1992 Oct;6(5):361–365. doi: 10.1016/0890-8508(92)90028-v. [DOI] [PubMed] [Google Scholar]
  7. Chatterton J. M., Joss A. W., Davidson M. M., Ho-Yen D. O. Why have Pneumocystis carinii trophozoites been ignored? J Clin Pathol. 1990 Apr;43(4):265–268. doi: 10.1136/jcp.43.4.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cushion M. T., Kaselis M., Stringer S. L., Stringer J. R. Genetic stability and diversity of Pneumocystis carinii infecting rat colonies. Infect Immun. 1993 Nov;61(11):4801–4813. doi: 10.1128/iai.61.11.4801-4813.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cushion M. T., Ruffolo J. J., Linke M. J., Walzer P. D. Pneumocystis carinii: growth variables and estimates in the A549 and WI-38 VA13 human cell lines. Exp Parasitol. 1985 Aug;60(1):43–54. doi: 10.1016/s0014-4894(85)80021-7. [DOI] [PubMed] [Google Scholar]
  10. Durkin M. M., Bartlett M. S., Queener S. F., Shaw M. M., Lee C. H., Smith J. W. An enzyme-linked immunosorbent assay for enumeration of Pneumocystis carinii in vitro and in vivo. J Clin Microbiol. 1992 Dec;30(12):3258–3262. doi: 10.1128/jcm.30.12.3258-3262.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Erdman S. M., Rodvold K. A. Trimethoprim/sulfamethoxazole concentrations in pneumocystis carinii Pneumonia. DICP. 1991 Sep;25(9):941–942. [PubMed] [Google Scholar]
  12. Freedberg K. A., Tosteson A. N., Cotton D. J., Goldman L. Optimal management strategies for HIV-infected patients who present with cough or dyspnea: a cost-effective analysis. J Gen Intern Med. 1992 May-Jun;7(3):261–272. doi: 10.1007/BF02598081. [DOI] [PubMed] [Google Scholar]
  13. Hardy W. D. Prophylaxis of AIDS-related opportunistic infections (OIs). Current status and future strategies. AIDS Clin Rev. 1991:145–180. [PubMed] [Google Scholar]
  14. Hughes W. T. Prevention and treatment of Pneumocystis carinii pneumonia. Annu Rev Med. 1991;42:287–295. doi: 10.1146/annurev.me.42.020191.001443. [DOI] [PubMed] [Google Scholar]
  15. Kim C. K., Foy J. M., Cushion M. T., Stanforth D., Linke M. J., Hendrix H. L., Walzer P. D. Comparison of histologic and quantitative techniques in evaluation of therapy for experimental Pneumocystis carinii pneumonia. Antimicrob Agents Chemother. 1987 Feb;31(2):197–201. doi: 10.1128/aac.31.2.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kitada K., Oka S., Kimura S., Shimada K., Serikawa T., Yamada J., Tsunoo H., Egawa K., Nakamura Y. Detection of Pneumocystis carinii sequences by polymerase chain reaction: animal models and clinical application to noninvasive specimens. J Clin Microbiol. 1991 Sep;29(9):1985–1990. doi: 10.1128/jcm.29.9.1985-1990.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liberator P. A., Anderson J. W., Powles M., Pittarelli L. A., Worley M., Becker-Hapak M., Graves D. C., Schmatz D. M. Comparative study of antipneumocystis agents in rats by using a Pneumocystis carinii-specific DNA probe to quantitate infection. J Clin Microbiol. 1992 Nov;30(11):2968–2974. doi: 10.1128/jcm.30.11.2968-2974.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lipschik G. Y., Gill V. J., Lundgren J. D., Andrawis V. A., Nelson N. A., Nielsen J. O., Ognibene F. P., Kovacs J. A. Improved diagnosis of Pneumocystis carinii infection by polymerase chain reaction on induced sputum and blood. Lancet. 1992 Jul 25;340(8813):203–206. doi: 10.1016/0140-6736(92)90469-j. [DOI] [PubMed] [Google Scholar]
  19. Lipschik G. Y., Masur H. Pneumocystis carinii pneumonia (PCP). Prog Clin Parasitol. 1991;2:27–71. [PubMed] [Google Scholar]
  20. Masur H. Prevention and treatment of pneumocystis pneumonia. N Engl J Med. 1992 Dec 24;327(26):1853–1860. doi: 10.1056/NEJM199212243272606. [DOI] [PubMed] [Google Scholar]
  21. Mulder J., McKinney N., Christopherson C., Sninsky J., Greenfield L., Kwok S. Rapid and simple PCR assay for quantitation of human immunodeficiency virus type 1 RNA in plasma: application to acute retroviral infection. J Clin Microbiol. 1994 Feb;32(2):292–300. doi: 10.1128/jcm.32.2.292-300.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Murry C. E., Schmidt R. A. Tissue invasion by Pneumocystis carinii: a possible cause of cavitary pneumonia and pneumothorax. Hum Pathol. 1992 Dec;23(12):1380–1387. doi: 10.1016/0046-8177(92)90058-b. [DOI] [PubMed] [Google Scholar]
  23. Olsson M., Elvin K., Löfdahl S., Linder E. Detection of Pneumocystis carinii DNA in sputum and bronchoalveolar lavage samples by polymerase chain reaction. J Clin Microbiol. 1993 Feb;31(2):221–226. doi: 10.1128/jcm.31.2.221-226.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Peten E. P., Striker L. J., Carome M. A., Elliott S. J., Yang C. W., Striker G. E. The contribution of increased collagen synthesis to human glomerulosclerosis: a quantitative analysis of alpha 2IV collagen mRNA expression by competitive polymerase chain reaction. J Exp Med. 1992 Dec 1;176(6):1571–1576. doi: 10.1084/jem.176.6.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Peters S. E., Wakefield A. E., Banerji S., Hopkin J. M. Quantification of the detection of Pneumocystis carinii by DNA amplification. Mol Cell Probes. 1992 Apr;6(2):115–117. doi: 10.1016/0890-8508(92)90055-3. [DOI] [PubMed] [Google Scholar]
  26. Peters S. E., Wakefield A. E., Sinclair K., Millard P. R., Hopkin J. M. A search for Pneumocystis carinii in post-mortem lungs by DNA amplification. J Pathol. 1992 Feb;166(2):195–198. doi: 10.1002/path.1711660217. [DOI] [PubMed] [Google Scholar]
  27. Schluger N., Godwin T., Sepkowitz K., Armstrong D., Bernard E., Rifkin M., Cerami A., Bucala R. Application of DNA amplification to pneumocystosis: presence of serum Pneumocystis carinii DNA during human and experimentally induced Pneumocystis carinii pneumonia. J Exp Med. 1992 Nov 1;176(5):1327–1333. doi: 10.1084/jem.176.5.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schreiber G., Dubeau L. C-myc proto-oncogene amplification detected by polymerase chain reaction in archival human ovarian carcinomas. Am J Pathol. 1990 Sep;137(3):653–658. [PMC free article] [PubMed] [Google Scholar]
  29. Sepkowitz K., Schluger N., Godwin T., Armstrong D., Cerami A., Bucala R. DNA amplification in experimental pneumocystosis: characterization of serum Pneumocystis carinii DNA and potential P. carinii carrier states. J Infect Dis. 1993 Aug;168(2):421–426. doi: 10.1093/infdis/168.2.421. [DOI] [PubMed] [Google Scholar]
  30. Thomas S., O'Doherty M., Bateman N. Pneumocystis carinii pneumonia. BMJ. 1990 Jan 27;300(6719):211–212. doi: 10.1136/bmj.300.6719.211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Travis W. D., Pittaluga S., Lipschik G. Y., Ognibene F. P., Suffredini A. F., Masur H., Feuerstein I., Kovacs J., Pass H. I., Condron K. S. Atypical pathologic manifestations of Pneumocystis carinii pneumonia in the acquired immune deficiency syndrome. Review of 123 lung biopsies from 76 patients with emphasis on cysts, vascular invasion, vasculitis, and granulomas. Am J Surg Pathol. 1990 Jul;14(7):615–625. doi: 10.1097/00000478-199007000-00002. [DOI] [PubMed] [Google Scholar]
  32. Wakefield A. E., Banerji S., Pixley F. J., Hopkin J. M. Molecular probes for the detection of Pneumocystis carinii. Trans R Soc Trop Med Hyg. 1990;84 (Suppl 1):17–18. doi: 10.1016/0035-9203(90)90450-s. [DOI] [PubMed] [Google Scholar]
  33. Wakefield A. E., Guiver L., Miller R. F., Hopkin J. M. DNA amplification on induced sputum samples for diagnosis of Pneumocystis carinii pneumonia. Lancet. 1991 Jun 8;337(8754):1378–1379. doi: 10.1016/0140-6736(91)93062-e. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Walzer P. D., Kim C. K., Foy J., Linke M. J., Cushion M. T. Cationic antitrypanosomal and other antimicrobial agents in the therapy of experimental Pneumocystis carinii pneumonia. Antimicrob Agents Chemother. 1988 Jun;32(6):896–905. doi: 10.1128/aac.32.6.896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Walzer P. D., LaBine M., Redington T. J., Cushion M. T. Lymphocyte changes during chronic administration of and withdrawal from corticosteroids: relation to Pneumocystis carinii pneumonia. J Immunol. 1984 Nov;133(5):2502–2508. [PubMed] [Google Scholar]
  37. Wong W. M., Lam V. M., Cheng L. Y., Tam J. W. Genomic sequence of a Sprague-Dawley rat beta-globin gene. Nucleic Acids Res. 1988 Mar 25;16(5):2342–2342. doi: 10.1093/nar/16.5.2342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wright C. F., Reid A. H., Tsai M. M., Ventre K. M., Murari P. J., Frizzera G., O'Leary T. J. Detection of Epstein-Barr virus sequences in Hodgkin's disease by the polymerase chain reaction. Am J Pathol. 1991 Aug;139(2):393–398. [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES