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. 2007 Nov 16;21(6):382–386. doi: 10.1002/jcla.20200

Flinders technology associates (FTA) filter paper–based DNA extraction with polymerase chain reaction (PCR) for detection of Pneumocystis jirovecii from respiratory specimens of immunocompromised patients

Surang Nuchprayoon 1,†,✉, Wilai Saksirisampant 1,‡, Siraya Jaijakul 1, Issarang Nuchprayoon 2
PMCID: PMC6649148  PMID: 18022923

Abstract

We evaluated the diagnostic value of Flinders Technology Associates (FTA) filter paper together with polymerase chain reaction (PCR) for detection of Pneumocystis jirovecii (carinii) from induced sputum (IS) and bronchoalveolar lavage fluid (BALF) samples. The study involved 162 patients with clinical diagnosis of pneumocystis pneumonia (PcP) of human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) patients and other immunocompromised patients. P. jirovecii cysts or trophozoites were detected in IS and BALF by cytological method. The mitochondrial 5S ribosomal ribonucleic acid (rRNA) gene of P. jirovecii was amplified from these samples by using FTA filters together with a one‐step PCR method (FTA‐PCR). With the FTA‐PCR method, the sensitivity and specificity of the test compared to microscopic examination were 67% and 90% for IS, while they were 67% and 91% for BALF, respectively. The sensitivity and specificity of the FTA‐PCR test was also comparable to PCR with the conventional deoxyribonucleic acid (DNA) extraction method. We concluded that FTA‐PCR is useful to detect P. jirovecii in noninvasive IS. J. Clin. Lab. Anal. 21:382–386, 2007. © 2007 Wiley‐Liss, Inc.

Keywords: Pneumocystis pneumonia, polymerase chain reaction, FTA filter paper, sputum, bronchoalveolar lavage fluid

REFERENCES

  • 1. Sritangratanakul S, Nuchprayoon S, Nuchprayoon I. Pneumocystis pneumonia: an update. J Med Assoc Thai 2004;87(Suppl 2):S309–S317. [PubMed] [Google Scholar]
  • 2. Stringer JR, Beard CB, Miller RF, Wakefield AE. A new name (Pneumocystis jiroveci) for Pneumocystis from humans. Emerg Infect Dis 2002;8:891–896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3. Frenkel JK. Pneumocystis jiroveci n. sp. from man: morphology, physiology, and immunology in relation to pathology. Natl Cancer Inst Monogr 1976;43:13–30. [PubMed] [Google Scholar]
  • 4. Edman JC, Kovacs JA, Masur H, Santi DV, Elwood HJ, Sogin ML. Ribosomal RNA sequence shows Pneumocystis carinii to be a member of the fungi. Nature 1988;334:519–522. [DOI] [PubMed] [Google Scholar]
  • 5. Gajdusek DC. Pneumocystis carinii; etiologic agent of interstitial plasma cell pneumonia of premature and young infants. Pediatrics 1957;19:543–565. [PubMed] [Google Scholar]
  • 6. Morris A, Beard CB, Huang L. Update on the epidemiology and transmission of Pneumocystis carinii . Microbes Infect 2002;4:95–103. [DOI] [PubMed] [Google Scholar]
  • 7. Hay JW, Osmond DH, Jacobson MA. Projecting the medical costs of AIDS and ARC in the United States. J Acquir Immune Defic Syndr 1988;1:466–485. [PubMed] [Google Scholar]
  • 8. Barlett MS, Smith JW. Pneumocystis carinii, an opportunist in immunocompromised patients. Clin Microbiol Rev 1991;4:137–149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. Masur H, Kaplan JE, Holmes KK. U.S. Public Health Service: infectious diseases society of America. Guidelines for preventing opportunistic infections among HIV‐infected persons—2002. Recommendations of the U.S. Public Health Service and the Infectious Diseases Society of America. Ann Intern Med 2002;137:435–478. [DOI] [PubMed] [Google Scholar]
  • 10. Lumbiganon P, Kosalaraksa P, Loapaiboon M. Survival of children with AIDS: experience in a university hospital in northeast Thailand. J Med Assoc Thai 2000;83:652–656. [PubMed] [Google Scholar]
  • 11. Saksirisampant W, Eampokalap B, Chantharodevong R, Changthong R. Comparison of methods for identification of Pneumocystis carinii in bronchoalveolar lavage fluid. J Med Assoc Thai 2002;85(Suppl 1):S407–S414. [PubMed] [Google Scholar]
  • 12. Wannamethee SG, Sirivichayakul S, Phillips AN, et al. Clinical and immunological features of human immunodeficiency virus infection in patients from Bangkok, Thailand. Int J Epidemiol 1998;27:289–295. [DOI] [PubMed] [Google Scholar]
  • 13. Torrington KG, Finelli MR. Small volume bronchoalveolar lavage used in diagnosing Pneumocystis carinii pneumonia in HIV‐infected patients. Chest 1995;107:1013–1017. [DOI] [PubMed] [Google Scholar]
  • 14. Jaijakul S, Saksirisampant W, Prownebon J, et al. Pneumocystis jiroveci in HIV/AIDS patients: detection by FTA filter paper together with PCR in noninvasive induced sputum specimens. J Med Assoc Thai 2005;88(Suppl 4):S294–S299. [PubMed] [Google Scholar]
  • 15. Orlandi PA, Lampel KA. Extraction‐free, filter‐based template preparation for rapid and sensitive PCR detection of pathogenic parasitic protozoa. J Clin Microbiol 2000;38:2271–2277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16. Subrungruang I, Mungthin M, Chavalitshewinkoon‐Petmitr P, Rangsin R, Naaglor T, Leelayoova S. Evaluation of DNA extraction and PCR methods for detection of Enterocytozoon bienuesi in stool specimens. J Clin Microbiol 2004;42:3490–3494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. Wakefield AE, Pixley FJ, Banerji S, et al. Detection of Pneumocystis carinii with DNA amplification. Lancet 1990;336:451–453. [DOI] [PubMed] [Google Scholar]
  • 18. Ruxrungtham K, Phanuphak P. Update on HIV/AIDS in Thailand. J Med Assoc Thai 2001;84(Suppl 1):S1–S17. [PubMed] [Google Scholar]
  • 19. Kim YK, Parulekar S, Yu PK, Pisani RJ, Smith TF, Anhalt JP. Evaluation of calcofluor white stain for detection of Pneumocystis carinii . Diagn Microbiol Infect Dis 1990;13:307–310. [DOI] [PubMed] [Google Scholar]
  • 20. Durand‐Joly I, Chabe M, Soula F, Delhaes L, Camus D, Dei‐Cas E. Molecular diagnosis of Pneumocystis pneumonia. FEMS Immunol Med Microbiol 2005;45:405–410. [DOI] [PubMed] [Google Scholar]
  • 21. Nuchprayoon S, Junpee A, Poovorawan Y, Scott AL. Detection and differentiation of filarial parasites by universal primers and polymerase chain reaction‐restriction fragment length polymerase analysis. Am J Trop Med Hyg 2005;73:895–900. [PubMed] [Google Scholar]
  • 22. Nuchprayoon S, Junpee A, Nithiuthai S, Chungpivat S, Suvannadabba S, Poovorawan Y. Detection of filarial parasites in domestic cats by PCR‐RFLP of ITS1. Vet Parasitol 2006;140:366–372. [DOI] [PubMed] [Google Scholar]
  • 23. Triteeraprapab S, Kanjanopas K, Suvannadabba S, Poovorawan Y, Scott AL. Transmission of the nocturnal periodic strain of Wuchereria bancriofti by Culex quinquefasciatus: establishing the potential for urban filariasis in Thailand. Epidemiol Infect 2000;125:207–212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. Ribes JA, Limper AH, Espy MJ, Smith TF. PCR detection of Pneumocystis carinii in bronchoalveolar lavage specimens: analysis of sensitivity and specificity. J Clin Microbiol 1997;35:830–835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. Sambrook J, Fritsch EF, Maniatis T. Preparation of reagents and buffers used in molecular cloning, p. B12 In: Sambrook J, Fritsch EF, Maniatis T, editors. Molecular cloning: a laboratory manual, 2nd edition New York: Cold Spring Harbor Laboratory Press; 1989. [Google Scholar]
  • 26. Mirdha BR, Guleria R. Comparative yield of different respiratory samples for diagnosis of Pneumocystis carinii infections in HIV‐seropositive and seronegative individuals in India. Southeast Asian J Trop Med Public Health 2000;31:473–477. [PubMed] [Google Scholar]
  • 27. Pinlaor S, Mootsikapun P, Pinlaor P, et al. PCR diagnosis of Pneumocystis carinii on sputum and bronchoalveolar lavage samples in immuno‐compromised patients. Parasitol Res 2004;94:213–218. [DOI] [PubMed] [Google Scholar]
  • 28. Rabodonirina M, Raffenot D, Cotte L, et al. Rapid detection of Pneumocystis carinii in bronchoalveolar lavage specimens from human immunodeficiency virus‐infected patients: use of a simple DNA extraction procedure and nested PCR. J Clin Microbiol 1997;35:2748–2751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29. Torres J, Goldman M, Wheat LJ, et al. Diagnosis of Pneumocystis carinii pneumonia in human immunodeficiency virus‐infected patients with polymerase chain reaction: a blinded comparison to standard methods. Clin Infect Dis 2000;30:141–145. [DOI] [PubMed] [Google Scholar]

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