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PCR diagnosis of Pneumocystis carinii on sputum and bronchoalveolar lavage samples in immuno-compromised patients

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Abstract

The polymerase chain reaction (PCR) technique was compared with Wright-Giemsa (WG), Gomori methenamine silver (GMS) stains and an immunofluorescence assay (IFA) for detection of Pneumocystis carinii in immuno-compromised patients. Specimens of 21 bronchoalveolar lavages (BAL) and 139 sputum samples, were obtained from 157 patients (38 with AIDS and 119 with HIV) from four hospitals in Khon Kaen, Thailand. A true positive required at least two positives by techniques considered gold standard tests. Eleven (52.38%) BAL and 13 (9.35%) sputum specimens were positive. PCR produced the highest sensitivity and negative predictive values for the BAL (100% for each) vs. sputum samples at 84.62 and 98.41 percent, respectively. The specificity of PCR was 90% and 98.41% for BAL and sputum samples, respectively. We suggest PCR is an important tool for the epidemiological study of P. carinii in high-risk individuals.

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References

  • Baughman RP, Liming JD (1998) Diagnostic strategies in Pneumocystis carinii pneumonia. Front Biosci 3:E1–E12

    CAS  PubMed  Google Scholar 

  • Caliendo AM, Hewitt PL, Allega JM, Keen A, Ruoff KL, Ferraro MJ (1998) Performance of a PCR assay for detection of Pneumocystis carinii from respiratory specimens. J Clin Microbiol 36:979–982

    CAS  PubMed  Google Scholar 

  • Cregan P, Yamamoto A, Lum A, VanDerHeide T, MacDonald M, Pulliam L (1990) Comparison of four methods for rapid detection of Pneumocystis carinii in respiratory specimens. J Clin Microbiol 28:2432–2436

    CAS  PubMed  Google Scholar 

  • Elvin K (1994) Laboratory diagnosis and occurrence of Pneumocystis carinii. Scand J Infect Dis Suppl 94:1–34

    CAS  PubMed  Google Scholar 

  • Hogan TF, Riley RS, Thomas JG (1997) Rapid diagnosis of acute eosinophilic pneumonia (AEP) in a patient with respiratory failure using bronchoalveolar lavage (BAL) with calcofluor white (CW) staining. J Clin Lab Anal 11:202–207

    Article  CAS  PubMed  Google Scholar 

  • Khan MA, Farrag N, Butcher P (1999) Diagnosis of Pneumocystis carinii pneumonia: immuno-fluorescence staining, simple PCR or nPCR. J Infect 39:77–80

    Article  CAS  PubMed  Google Scholar 

  • Kitada K, Oka S, Kohjin T, Kimura S, Nakamura Y, Shimada K (1993) Pneumocystis carinii pneumonia monitored by P. carinii shedding in sputum by the polymerase chain reaction. Intern Med 32:370–373

    CAS  PubMed  Google Scholar 

  • Kovacs JA, Gill V, Swan JC, Ognibene F, Shelhamer J, Parrillo JE, Masur H (1986) Prospective evaluation of a monoclonal antibody in diagnosis of Pneumocysti carinii pneumonia. Lancet 2:1–3

    Article  CAS  PubMed  Google Scholar 

  • Kovacs JA, Ng VL, Masur H, Leoung G, Hadley WK, Evans G, Lane HC, Ognibene FP, Shelhamer J, Parrillo JE et al. (1988) Diagnosis of Pneumocystis carinii pneumonia: improved detection in sputum with use of monoclonal antibodies. N Engl J Med 318:589–593

    CAS  PubMed  Google Scholar 

  • Larsen HH, Masur H, Kovacs JA, Gill VJ, Silcott VA, Kogulan P, Maenza J, Smith M, Lucey DR, Fischer SH (2002) Development and evaluation of a quantitative, touch-down, real-time PCR assay for diagnosing Pneumocystis carinii pneumonia. J Clin Microbiol 40:490–494

    Article  CAS  PubMed  Google Scholar 

  • Lasbury ME, Lin P, Tschang D, Durant PJ, Lee CH (2004) Effect of bronchoalveolar lavage fluid from Pneumocystis carinii- infected hosts on phagocytic activity of alveolar macrophages. Infect Immun 72:2140–2147

    Google Scholar 

  • Leibovitz E, Pollack H, Moore T, Papellas J, Gallo L, Krasinski K, Borkowsky W (1995) Comparison of PCR and standard cytological staining for detection of Pneumocystis carinii from respiratory specimens from patients with or at high risk for infection by human immunodeficiency virus. J Clin Microbiol 33:3004–3007

    CAS  PubMed  Google Scholar 

  • Lu JJ, Chen CH, Bartlett MS, Smith JW, Lee CH (1995) Comparison of six different PCR methods for detection of Pneumocystis carinii. J Clin Microbiol 33:2785–2788

    CAS  PubMed  Google Scholar 

  • Ng VL, Virani NA, Chaisson RE, Yajko DM, Sphar HT, Cabrian K, Rollins N, Charache P, Krieger M, Hadley WK, Hopewell PC (1990) Rapid detection of Pneumocystis carinii using a direct fluorescent monoclonal antibody stain. J Clin Microbiol 28:2228–2233

    CAS  PubMed  Google Scholar 

  • Olsson M, Elvin K, Lofdahl S, Linder E (1993) Detection of Pneumocystis carinii DNA in sputum and bronchoalveolar lavage samples by polymerase chain reaction. J Clin Microbiol 31:221–226

    CAS  PubMed  Google Scholar 

  • Ruffini DD, Madhi SA (2002) The high burden of Pneumocystis carinii pneumonia in African HIV-1-infected children hospitalized for severe pneumonia. Aids 15 16:105–112

    Article  Google Scholar 

  • Ruxrungtham K, Phanuphak P (2001) Update on HIV/AIDS in Thailand. J Med Assoc Thai 84 Suppl 1:S1–17

    Google Scholar 

  • Saksirisampant W, Eampokalap B, Chantharodevong R, Changthong R (2002) Comparison of methods for identification of Pneumocystis carinii in bronchoalveolar lavage fluid. J Med Assoc Thai 85 Suppl 1:S407–414

    Google Scholar 

  • Sandhu GS, Kline BC, Espy MJ, Stockman L, Smith TF, Limper AH (1999) Laboratory diagnosis of Pneumocystis carinii infections by PCR directed to genes encoding for mitochondrial 5S and 28S ribosomal RNA. Diagn Microbiol Infect Dis 33:157–162

    Article  CAS  PubMed  Google Scholar 

  • Shiota T (1986) Simultaneous demonstration of cyst walls and intracystic bodies of Pneumocystis carinii in paraffin embedded lung sections using Gomori’s methenamine silver nitrate and Giemsa stain. J Clin Pathol 39:1269–1271

    CAS  PubMed  Google Scholar 

  • Tollerud DJ, Wesseler TA, Kim CK, Baughman RP (1989) Use of a rapid differential stain for identifying Pneumocystis carinii in bronchoalveolar lavage fluid. Diagnostic efficacy in patients with AIDS. Chest 95:494–497

    CAS  PubMed  Google Scholar 

  • Tuncer S, Erguven S, Kocagoz S, Unal S (1998) Comparison of cytochemical staining, immuno-fluorescence and PCR for diagnosis of Pneumocystis carinii on sputum samples. Scand J Infect Dis 30:125–128

    Article  CAS  PubMed  Google Scholar 

  • Turner D, Schwarz Y, Yust I (2003) Induced sputum for diagnosing Pneumocystis carinii pneumonia in HIV patients: new data, new issues. Eur Respir J 21:204–208

    Article  CAS  PubMed  Google Scholar 

  • Wakefield AE (2002) Pneumocystis carinii. Br Med Bull 61:175–188

    Article  CAS  PubMed  Google Scholar 

  • Wakefield AE, Guiver L, Miller RF, Hopkin JM (1991) DNA amplification on induced sputum samples for diagnosis of Pneumocystis carinii pneumonia. Lancet 337:1378–1379

    Article  CAS  PubMed  Google Scholar 

  • Weig M, Klinker H, Wilhelm M, Lemmer K, Gross U (1996) Correlation of Pneumocystis carinii PCR with clinical diagnosis in immuno-compromised patients. Lancet 347:1266

    Article  CAS  Google Scholar 

  • Wilkin A, Feinberg J (1999) Pneumocystis carinii pneumonia: a clinical review. Am Fam Physician 60:1699–1708, 1713–1714

    CAS  PubMed  Google Scholar 

  • Yoshida EM, Filipenko D, Phillips P, Montaner JS, Whittaker JS (1996) AIDS-related extrapulmonary Pneumocystis carinii infection presenting as a solitary rectal ulcer. Can J Gastroenterol 10:401–404

    CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the Ministry of University Affairs, Thailand. We thank all the nurses and physicians for assisting in the gathering of specimens, Dr. Nongluk Tesana, a physician at the Tuberculosis Center, Khon Kaen, Thailand, for prepping the patients, and Mr. Bryan Roderick Hamman for assistance with the English-language presentation. We are also very grateful to Professor James A. Will for improving the scientific English-language presentation.

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Authors and Affiliations

  1. Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand

    Somchai Pinlaor, Vichit Pipitgool & Paiboon Sithithaworn

  2. Department of Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand

    Piroon Mootsikapun & Anakapong Phunmanee

  3. Department of Clinical Microbiology, Faculty of Associated Medical Science, Khon Kaen University, Khon Kaen 40002, Thailand

    Porntip Pinlaor

  4. Department of Clinical Microscopy, Faculty of Associated Medical Science, Khon Kaen University, Khon Kaen 40002, Thailand

    Worawan Chumpia & Jiraporn Sithithaworn

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  1. Somchai Pinlaor
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  2. Piroon Mootsikapun
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  5. Vichit Pipitgool
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  8. Jiraporn Sithithaworn
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Correspondence to Somchai Pinlaor.

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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 94, 213–218 (2004). https://doi.org/10.1007/s00436-004-1200-y

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  • DOI: https://doi.org/10.1007/s00436-004-1200-y

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