1887

Abstract

pneumonia (PCP) is an opportunistic infection affecting immunocompromised patients. While conventional diagnosis of PCP by microscopy is cumbersome, the use of PCR to diagnose PCP has great potential. Nevertheless, inter-laboratory validation and standardization of PCR assays is lacking. The aim of this study was to evaluate the inter-laboratory agreement of three independently developed real-time PCR assays for the detection of in bronchoalveolar lavage fluid samples. Therefore, 124 samples were collected in three tertiary care laboratories (Leiden University Medical Center, Maastricht Infection Center and Radboud University Nijmegen Medical Centre) and were tested by both microscopy and real-time PCR. Of 41 samples positive for by microscopy, 40 were positive in all three PCR assays. The remaining sample was positive in a single assay only. Out of 83 microscopy-negative samples, 69 were negative in all three PCR assays. The other 14 samples were found positive, either in all three assays (=5), in two (=2) or in one of the assays (=7). The data demonstrate high inter-laboratory agreement among real-time PCR assays for the detection of .

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.46552-0
2006-09-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jmm/55/9/1229.html?itemId=/content/journal/jmm/10.1099/jmm.0.46552-0&mimeType=html&fmt=ahah

References

  1. Amin M. B., Mezger E., Zarbo R. J. 1992; Detection of Pneumocystis carinii . Comparative study of monoclonal antibody and silver staining. Am J Clin Pathol 98:13–18
    [Google Scholar]
  2. Chandra P., Delaney M. D., Tuazon C. U. 1988; Role of special stains in the diagnosis of Pneumocystis carinii infection from bronchial washing specimens in patients with the acquired immune deficiency syndrome. Acta Cytol 32:105–108
    [Google Scholar]
  3. Djamin R. S., Drent M., Schreurs A. J., Groen E. A., Wagenaar S. S. 1998; Diagnosis of Pneumocystis carinii pneumonia in HIV-positive patients. Bronchoalveolar lavage vs. bronchial brushing. Acta Cytol 42:933–938 [CrossRef]
    [Google Scholar]
  4. Edman J. C., Kovacs J. A., Masur H., Santi D., Elwood V. H. J., Sogin M. L. 1988; Ribosomal RNA sequence shows Pneumocystis carinii to be a member of the fungi. Nature 334:519–522 [CrossRef]
    [Google Scholar]
  5. Elvin K. M., Bjorkman A., Linder E., Heurlin N., Hjerpe A. 1988; Pneumocystis carinii pneumonia: detection of parasites in sputum and bronchoalveolar lavage fluid by monoclonal antibodies. BMJ 297:381–384 [CrossRef]
    [Google Scholar]
  6. Flori P., Bellete B., Durand F., Raberin H., Cazorla C., Hafid J., Lucht F., Sung R. T. 2004; Comparison between real-time PCR, conventional PCR and different staining techniques for diagnosing Pneumocystis jiroveci pneumonia from bronchoalveolar lavage specimens. J Med Microbiol 53:603–607 [CrossRef]
    [Google Scholar]
  7. Jacobs J. A., Dieleman M. M., Cornelissen E. I., Groen E. A., Wagenaar S. S., Drent M. 2001; Bronchoalveolar lavage fluid cytology in patients with Pneumocystis carinii pneumonia. Acta Cytol 45317–326
    [Google Scholar]
  8. Kovacs J. A., Ng V. L., Masur H. & 7 other authors; 1988; Diagnosis of Pneumocystis carinii pneumonia: improved detection in sputum with use of monoclonal antibodies. N Engl J Med 318:589–593 [CrossRef]
    [Google Scholar]
  9. Kovacs J. A., Gill V. J., Meshnick S., Masur H. 2001; New insights into transmission, diagnosis, and drug treatment of Pneumocystis carinii pneumonia. JAMA (J Am Med Assoc) 286:2450–2460 [CrossRef]
    [Google Scholar]
  10. Larsen H. H., Kovacs J. A. F., Stock F., Vestereng V. H., Lundgren B., Fischer S. H., Gill V. J. 2002a; Development of a rapid real-time PCR assay for quantitation of Pneumocystis carinii f. sp. carinii . J Clin Microbiol 40:2989–2993 [CrossRef]
    [Google Scholar]
  11. Larsen H. H., Masur H., Kovacs J. A. & 7 other authors; 2002b; Development and evaluation of a quantitative, touch-down, real-time PCR assay for diagnosing Pneumocystis carinii pneumonia. J Clin Microbiol 40:490–494 [CrossRef]
    [Google Scholar]
  12. Larsen H. H., Huang L., Kovacs J. A. & 7 other authors; 2004; A prospective, blinded study of quantitative touch-down polymerase chain reaction using oral-wash samples for diagnosis of Pneumocystis pneumonia in HIV-infected patients. J Infect Dis 189:1679–1683 [CrossRef]
    [Google Scholar]
  13. Limper A. H., Offord K. P., Smith T. F., Martin W. J. 2nd (1989; Pneumocystis carinii pneumonia . Differences in lung parasite number and inflammation in patients with and without AIDS. Am Rev Respir Dis 140:1204–1209 [CrossRef]
    [Google Scholar]
  14. Maskell N. A., Waine D. J., Lindley A., Pepperell J. C., Wakefield A. E., Miller R. F., Davies R. J. 2003; Asymptomatic carriage of Pneumocystis jiroveci in subjects undergoing bronchoscopy: a prospective study. Thorax 58:594–597 [CrossRef]
    [Google Scholar]
  15. Miller R. F. 1999; Pneumocystis carinii infection in non-AIDS patients. Curr Opin Infect Dis 12:371–377 [CrossRef]
    [Google Scholar]
  16. Miller R. F., Ambrose H. E., Wakefield A. E. 2001; Pneumocystis carinii f. sp. hominis DNA in immunocompetent health care workers in contact with patients with P. carinii pneumonia. J Clin Microbiol 39:3877–3882 [CrossRef]
    [Google Scholar]
  17. Nevez G., Guyot K., Totet A., Raccurt C., Dei-Cas C. 2001; Pulmonary colonisation with Pneumocystis carinii in an immunosuppressed HIV-negative patient: detection and typing of the fungus by PCR. J Med Microbiol 50:198–200
    [Google Scholar]
  18. Olsson M., Stralin K., Holmberg H. 2001; Clinical significance of nested polymerase chain reaction and immunofluorescence for detection of Pneumocystis carinii pneumonia. Clin Microbiol Infect 7:492–497 [CrossRef]
    [Google Scholar]
  19. Sepkowitz K. A. 2002; Opportunistic infections in patients with and patients without Acquired Immunodeficiency Syndrome. Clin Infect Dis 34:1098–1107 [CrossRef]
    [Google Scholar]
  20. Sing A., Roggenkamp A., Autenrieth I. B., Heesemann J. 1999; Pneumocystis carinii carriage in immunocompetent patients with primary pulmonary disorders as detected by single or nested PCR. J Clin Microbiol 37:3409–3410
    [Google Scholar]
  21. Stringer J. R., Cushion M. T., Wakefield A. E. 2001; New nomenclature for the genus Pneumocystis. J Eukaryot Microbiol Suppl184S–189S
    [Google Scholar]
  22. Thomas C. F. Jr, Limper A. H. 2004; Pneumocystis pneumonia . N Engl J Med 350:2487–2498 [CrossRef]
    [Google Scholar]
  23. Visconti E., Marinaci S., Zolfo M., Mencarini P., Tamburrini E., Pagliari G., Ortona E., Siracusano A. 2000; Very low frequence of Pneumocystis carinii DNA detection by PCR in specimens from patients with lung damage. J Clin Microbiol 38:1307–1308
    [Google Scholar]
  24. Wakefield A. E., Pixley F. J., Banerji S., Sinclair K., Miller R. F., Moxon E. R., Hopkin J. M. 1990; Detection of Pneumocystis carinii with DNA amplification. Lancet 336:451–453 [CrossRef]
    [Google Scholar]
  25. Weig M., Klinker H., Wilhelm M., Lemmer K., Gross U. 1996; Correlation of Pneumocystis carinii PCR with clinical diagnosis in immunocompromised patients. Lancet 347:1266
    [Google Scholar]
  26. Weig M., Klinker H., Bogner B. H., Meier A., Gross U. 1997; Usefulness of PCR for diagnosis of Pneumocystis carinii pneumonia in different patient groups. J Clin Microbiol 35:1445–1449
    [Google Scholar]
  27. Yale S. H., Limper A. H. 1996; Pneumocystis carinii pneumonia in patients without acquired immunodeficiency syndrome: associated illness and prior corticosteroid therapy. Mayo Clin Proc 71:5–13 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.46552-0
Loading
/content/journal/jmm/10.1099/jmm.0.46552-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error