Abstract
The diagnosis of invasive pulmonary aspergillosis (IPA) is still in challenge in clinical practice, particularly for those patients without an obvious neutropaenia. In this study, a well-validated qPCR method and Platelia galactomannan (GM) assay were compared for their diagnostic performance using paired samples of bronchoalveolar lavage (BAL) fluid and serum from predominantly non-neutropaenic patients. In the serum samples, qPCR showed a comparable performance with GM assay in terms of sensitivity and specificity. In the BAL samples, qPCR and GM assay both demonstrated a good sensitivity (90 vs. 90 %); however, the specificity of qPCR was higher than that of GM assay (92.5 vs. 68.8 %, P < 0.001) in these samples. A better sensitivity was obtained with BAL compared with serum samples for both GM assay (90 vs. 50 %) and qPCR (90 vs. 60 %). In conclusion, in non-neutropaenic patients, BAL appears to provide improved sensitivity for both GM and qPCR assays. BAL qPCR offers a better diagnostic value for IPA compared with BAL GM assay, significantly increasing the specificity without affecting the sensitivity.
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Kontoyiannis DP, Marr KA, Park BJ, et al. Prospective surveillance for invasive fungal infections in hematopoietic stem cell transplant recipients, 2001–2006: overview of the transplant-associated infection surveillance network (TRANSNET) database. Clin Infect Dis. 2010;50:1091–100.
Pagano L, Caira M, Candoni A, et al. Invasive aspergillosis in patients with acute myeloid leukemia: a SEIFEM-2008 registry study. Haematologica. 2010;95:644–50.
Guinea J, Torres-Narbona M, Gijón P, et al. Pulmonary aspergillosis in patients with chronic obstructive pulmonary disease: incidence, risk factors, and outcome. Clin Microbiol Infect. 2010;16:870–7.
Trof RJ, Beishuizen A, Debets-Ossenkopp YJ, Girbes AR, Groeneveld AB. Management of invasive pulmonary aspergillosis in non-neutropenic critically ill patients. Intensive Care Med. 2007;33:1694–703.
Dai Z, Zhao H, Cai S, Lv Y, Tong W. Invasive pulmonary aspergillosis in non-neutropenic patients with and without underlying disease: a single-centre retrospective analysis of 52 subjects. Respirology. 2013;18:323–31.
Ostrosky-Zeichner L. Invasive mycoses: diagnostic challenges. Am J Med. 2012;125(1 Suppl):S14–24.
Hope WW, Walsh TJ, Denning DW. Laboratory diagnosis of invasive aspergillosis. Lancet Infect Dis. 2005;5:609–22.
Tarrand JJ, Lichterfeld M, Warraich I, et al. Diagnosis of invasive septate mold infections. A correlation of microbiological culture and histologic or cytologic examination. Am J Clin Pathol. 2003;119:854–8.
Greene RE, Schlamm HT, Oestmann JW, et al. Imaging findings in acute invasive pulmonary aspergillosis: clinical significance of the halo sign. Clin Infect Dis. 2007;44:373–9.
Bergeron A, Porcher R, Sulahian A, et al. The strategy for the diagnosis of invasive pulmonary aspergillosis should depend on both the underlying condition and the leukocyte count of patients with hematologic malignancies. Blood. 2012;119:1831–7 (quiz 1956).
Zhang R, Wang S, Lu H, Wang Z, Xu X. Misdiagnosis of invasive pulmonary aspergillosis: a clinical analysis of 26 immunocompetent patients. Int J Clin Exp Med. 2014;7:5075–82.
Pfeiffer CD, Fine JP, Safdar N. Diagnosis of invasive aspergillosis using a galactomannan assay: a meta-analysis. Clin Infect Dis. 2006;42:1417–27.
Cai X, Ni W, Wei C, Cui J. Diagnostic value of the serum galactomannan and (1,3)-beta-d-glucan assays for invasive pulmonary aspergillosis in non-neutropenic patients. Intern Med. 2014;53:2433–7.
Teering S, Verreth A, Peeters A, et al. Prognostic value of serum galactomannan in mixed ICU patients: a retrospective observational study. Anaesthesiol Intensive Ther. 2014;46:145–54.
Mengoli C, Cruciani M, Barnes RA, Loeffler J, Donnelly JP. Use of PCR for diagnosis of invasive aspergillosis: systematic review and meta-analysis. Lancet Infect Dis. 2009;9:89–96.
Luong ML, Clancy CJ, Vadnerkar A, et al. Comparison of an aspergillus real-time polymerase chain reaction assay with galactomannan testing of bronchoalvelolar lavage fluid for the diagnosis of invasive pulmonary aspergillosis in lung transplant recipients. Clin Infect Dis. 2011;52:1218–26.
Challier S, Boyer S, Abachin E, Berche P. Development of a serum-based Taqman real-time PCR assay for diagnosis of invasive aspergillosis. J Clin Microbiol. 2004;42:844–6.
White PL, Linton CJ, Perry MD, Johnson EM, Barnes RA. The evolution and evaluation of a whole blood polymerase chain reaction assay for the detection of invasive aspergillosis in hematology patients in a routine clinical setting. Clin Infect Dis. 2006;42:479–86.
White PL, Perry MD, Moody A, Follett SA, Morgan G, Barnes RA. Evaluation of analytical and preliminary clinical performance of Myconostica MycAssay Aspergillus when testing serum specimens for diagnosis of invasive Aspergillosis. J Clin Microbiol. 2011;49:2169–74.
Torelli R, Sanguinetti M, Moody A, et al. Diagnosis of invasive aspergillosis by a commercial real-time PCR assay for aspergillus DNA in bronchoalveolar lavage fluid samples from high-risk patients compared to a galactomannan enzyme immunoassay. J Clin Microbiol. 2011;49:4273–8.
Sanguinetti M, Posteraro B, Pagano L, et al. Comparison of real-time PCR, conventional PCR, and galactomannan antigen detection by enzyme-linked immunosorbent assay using bronchoalveolar lavage fluid samples from hematology patients for diagnosis of invasive pulmonary aspergillosis. J Clin Microbiol. 2003;41:3922–5.
Khot PD, Ko DL, Hackman RC, Fredricks DN. Development and optimization of quantitative PCR for the diagnosis of invasive aspergillosis with bronchoalveolar lavage fluid. BMC Infect Dis. 2008;8:73.
Bernal-Martinez L, Gago S, Buitrago MJ, Gomez-Lopez A, Rodriguez-Tudela JL, Cuenca-Estrella M. Analysis of performance of a PCR-based assay to detect DNA of Aspergillus fumigatus in whole blood and serum: a comparative study with clinical samples. J Clin Microbiol. 2011;49:3596–9.
Spiess B, Buchheidt D, Baust C, et al. Development of a LightCycler PCR assay for detection and quantification of Aspergillus fumigatus DNA in clinical samples from neutropenic patients. J Clin Microbiol. 2003;41:1811–8.
Guinea J, Padilla C, Escribano P, et al. Evaluation of MycAssay aspergillus for diagnosis of invasive pulmonary aspergillosis in patients without hematological cancer. PLoS ONE. 2013;8:e61545.
De Pauw B, Walsh TJ, Donnelly JP, et al. Revised definitions of invasive fungal disease from the European organization for research and treatment of cancer/invasive fungal infections cooperative group and the national institute of allergy and infectious diseases mycoses study group (EORTC/MSG) consensus group. Clin Infect Dis. 2008;46:1813–21.
Hanley JA, McNeil BJ. The meaning and use of the area under a receiver operating characteristic (ROC) curve. Radiology. 1982;143:29–36.
Pini P, Bettua C, Orsi CF, et al. Clinical performance of a commercial real-time PCR assay for aspergillus DNA detection in serum samples from high-risk patients: comparison with a galactomannan enzyme immunoassay. Eur J Clin Microbiol Infect Dis. 2014. doi:10.1007/s10096-014-2211-y.
Lewis RE, Kontoyiannis DP. Invasive aspergillosis in glucocorticoid-treated patients. Med Mycol. 2009;47(Suppl 1):S271–81.
Adam O, Auperin A, Wilquin F, Bourhis JH, Gachot B, Chachaty E. Treatment with piperacillin-tazobactam and false-positive aspergillus galactomannan antigen test results for patients with hematological malignancies. Clin Infect Dis. 2004;38:917–20.
Zhang S, Wang S, Wan Z, Li R, Yu J. The diagnosis of invasive and noninvasive pulmonary aspergillosis by serum and bronchoalveolar lavage fluid galactomannan assay. Biomed Res Int. 2015;2015:943691.
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This work was supported by National Science and Technology Major Project (2013ZX10004612-002).
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Zhang, S., Wang, S., Wan, Z. et al. Quantitative Real-Time PCR and Platelia Galactomannan Assay for the Diagnosis of Invasive Pulmonary Aspergillosis: Bronchoalveolar Lavage Fluid Performs Better Than Serum in Non-neutropaenic Patients. Mycopathologia 181, 625–629 (2016). https://doi.org/10.1007/s11046-016-0024-5
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DOI: https://doi.org/10.1007/s11046-016-0024-5