Jundishapur Journal of Microbiology

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Investigating the Presence of Aspergillus fumigatus and A. flavus Using Galactomannan Enzyme Assay and TaqMan Real-Time PCR Technique

Shahla Roudbarmohammadi 1 , Alireza Salimi-Khorashad 2 , * , Mehdi Forouzandeh-Moghadam 3 and Maryam Roudbary 4
Authors Information
1 Department of Medical Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2 Infectious Diseases and Tropical Medicine Research Center, Resistant Tuberculosis Institute, Zahedan University of Medical Sciences, Zahedan, Iran
3 Department of Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
4 Department of Medical Mycology and Parasitology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
Article information
  • Jundishapur Journal of Microbiology: February 2018, 11 (2); e13670
  • Published Online: February 10, 2018
  • Article Type: Research Article
  • Received: May 23, 2017
  • Revised: November 28, 2017
  • Accepted: December 9, 2017
  • DOI: 10.5812/jjm.13670

To Cite: Roudbarmohammadi S, Salimi-Khorashad A, Forouzandeh-Moghadam M, Roudbary M. Investigating the Presence of Aspergillus fumigatus and A. flavus Using Galactomannan Enzyme Assay and TaqMan Real-Time PCR Technique, Jundishapur J Microbiol. 2018 ; 11(2):e13670. doi: 10.5812/jjm.13670.

Copyright © 2018, Jundishapur Journal of Microbiology. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
1. Background
2. Objectives
3. Methods
4. Results
5. Discussion
6. Conclusions
  • 1. Latge JP. Aspergillus fumigatus and aspergillosis. Clin Microbiol Rev. 1999;12(2):310-50. [PubMed: 10194462].
  • 2. Ali R, Ozkalemkas F, Ozcelik T, Ozkocaman V, Ozkan A, Bayram S, et al. Invasive pulmonary aspergillosis: role of early diagnosis and surgical treatment in patients with acute leukemia. Ann Clin Microbiol Antimicrob. 2006;5:17. doi: 10.1186/1476-0711-5-17. [PubMed: 16872530].
  • 3. de Aguirre L, Hurst SF, Choi JS, Shin JH, Hinrikson HP, Morrison CJ. Rapid differentiation of Aspergillus species from other medically important opportunistic molds and yeasts by PCR-enzyme immunoassay. J Clin Microbiol. 2004;42(8):3495-504. doi: 10.1128/JCM.42.8.3495-3504.2004. [PubMed: 15297489].
  • 4. Hedayati MT, Mohseni-Bandpi A, Moradi S. A survey on the pathogenic fungi in soil samples of potted plants from Sari hospitals, Iran. J Hosp Infect. 2004;58(1):59-62. doi: 10.1016/j.jhin.2004.04.011. [PubMed: 15350715].
  • 5. Ferns RB. Evaluation of the role of real-time PCR in the diagnosis of invasive aspergillosis. Leuk Lymphoma. 2006;47(1):15-20. doi: 10.1080/10428190500244233. [PubMed: 16321822].
  • 6. Hayette MP, Vaira D, Susin F, Boland P, Christiaens G, Melin P, et al. Detection of Aspergillus species DNA by PCR in bronchoalveolar lavage fluid. J Clin Microbiol. 2001;39(6):2338-40. doi: 10.1128/JCM.39.6.2338-2340.2001. [PubMed: 11376086].
  • 7. Sanguinetti M, Posteraro B, Pagano L, Pagliari G, Fianchi L, Mele 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(8):3922-5. [PubMed: 12904419].
  • 8. Lee FJ, Lin LW, Smith JA. Purification and characterization of an N alpha-acetyltransferase from Saccharomyces cerevisiae. J Biol Chem. 1988;263(29):14948-55. [PubMed: 2844792].
  • 9. Zhang YJ, Zhang S, Liu XZ, Wen HA, Wang M. A simple method of genomic DNA extraction suitable for analysis of bulk fungal strains. Lett Appl Microbiol. 2010;51(1):114-8. doi: 10.1111/j.1472-765X.2010.02867.x. [PubMed: 20536704].
  • 10. Luong ML, Clancy CJ, Vadnerkar A, Kwak EJ, Silveira FP, Wissel MC, 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(10):1218-26. doi: 10.1093/cid/cir185. [PubMed: 21507918].
  • 11. Walsh TJ, Wissel MC, Grantham KJ, Petraitiene R, Petraitis V, Kasai M, et al. Molecular detection and species-specific identification of medically important Aspergillus species by real-time PCR in experimental invasive pulmonary aspergillosis. J Clin Microbiol. 2011;49(12):4150-7. doi: 10.1128/JCM.00570-11. [PubMed: 21976757].
  • 12. Becker MJ, Lugtenburg EJ, Cornelissen JJ, Van Der Schee C, Hoogsteden HC, De Marie S. Galactomannan detection in computerized tomography-based broncho-alveolar lavage fluid and serum in haematological patients at risk for invasive pulmonary aspergillosis. Br J Haematol. 2003;121(3):448-57. doi: 10.1046/j.1365-2141.2003.04308.x. [PubMed: 12716367].
  • 13. Verweij PE, Latge JP, Rijs AJ, Melchers WJ, De Pauw BE, Hoogkamp-Korstanje JA, et al. Comparison of antigen detection and PCR assay using bronchoalveolar lavage fluid for diagnosing invasive pulmonary aspergillosis in patients receiving treatment for hematological malignancies. J Clin Microbiol. 1995;33(12):3150-3. [PubMed: 8586691].
  • 14. Francesconi A, Kasai M, Petraitiene R, Petraitis V, Kelaher AM, Schaufele R, et al. Characterization and comparison of galactomannan enzyme immunoassay and quantitative real-time PCR assay for detection of Aspergillus fumigatus in bronchoalveolar lavage fluid from experimental invasive pulmonary aspergillosis. J Clin Microbiol. 2006;44(7):2475-80. doi: 10.1128/JCM.02693-05. [PubMed: 16825367].
  • 15. Hedayati MT, Khodavaisy S, Alialy M, Omran SM, Habibi MR. Invasive aspergillosis in intensive care unit patients in Iran. Acta Medica (Hradec Kralove). 2013;56(2):52-6. doi: 10.14712/18059694.2014.24. [PubMed: 24069658].
  • 16. Badiee P, Moghadami M, Rozbehani H. Comparing immunological and molecular tests with conventional methods in diagnosis of acute invasive fungal rhinosinusitis. J Infect Dev Ctries. 2016;10(1):90-5. doi: 10.3855/jidc.6411. [PubMed: 26829542].
  • 17. Barton RC. Laboratory diagnosis of invasive aspergillosis: From diagnosis to prediction of outcome. Scientifica. 2013;2013:1-29. doi: 10.1155/2013/459405.
  • 18. Becker MJ, de Marie S, Willemse D, Verbrugh HA, Bakker-Woudenberg IA. Quantitative galactomannan detection is superior to PCR in diagnosing and monitoring invasive pulmonary aspergillosis in an experimental rat model. J Clin Microbiol. 2000;38(4):1434-8. [PubMed: 10747121].
  • 19. Marr KA, Balajee SA, McLaughlin L, Tabouret M, Bentsen C, Walsh TJ. Detection of galactomannan antigenemia by enzyme immunoassay for the diagnosis of invasive aspergillosis: variables that affect performance. J Infect Dis. 2004;190(3):641-9. doi: 10.1086/422009. [PubMed: 15243943].
  • 20. Marr KA, Laverdiere M, Gugel A, Leisenring W. Antifungal therapy decreases sensitivity of the Aspergillus galactomannan enzyme immunoassay. Clin Infect Dis. 2005;40(12):1762-9. doi: 10.1086/429921. [PubMed: 15909264].
  • 21. Ansorg R, van den Boom R, Rath PM. Detection of Aspergillus galactomannan antigen in foods and antibiotics. Mycoses. 1997;40(9-10):353-7. doi: 10.1111/j.1439-0507.1997.tb00249.x. [PubMed: 9470421].
  • 22. Mattei D, Rapezzi D, Mordini N, Cuda F, Lo Nigro C, Musso M, et al. False-positive Aspergillus galactomannan enzyme-linked immunosorbent assay results in vivo during amoxicillin-clavulanic acid treatment. J Clin Microbiol. 2004;42(11):5362-3. doi: 10.1128/JCM.42.11.5362-5363.2004. [PubMed: 15528743].
  • 23. Aubry A, Porcher R, Bottero J, Touratier S, Leblanc T, Brethon B, et al. Occurrence and kinetics of false-positive Aspergillus galactomannan test results following treatment with beta-lactam antibiotics in patients with hematological disorders. J Clin Microbiol. 2006;44(2):389-94. doi: 10.1128/JCM.44.2.389-394.2006. [PubMed: 16455889].
  • 24. Sulahian A, Touratier S, Ribaud P. False positive test for aspergillus antigenemia related to concomitant administration of piperacillin and tazobactam. N Engl J Med. 2003;349(24):2366-7. doi: 10.1056/NEJM200312113492424. [PubMed: 14668472].
  • 25. Viscoli C, Machetti M, Cappellano P, Bucci B, Bruzzi P, Van Lint MT, et al. False-positive galactomannan platelia Aspergillus test results for patients receiving piperacillin-tazobactam. Clin Infect Dis. 2004;38(6):913-6. doi: 10.1086/382224. [PubMed: 14999640].
  • 26. Mikulska M, Furfaro E, Del Bono V, Raiola AM, Ratto S, Bacigalupo A, et al. Piperacillin/tazobactam (Tazocin) seems to be no longer responsible for false-positive results of the galactomannan assay. J Antimicrob Chemother. 2012;67(7):1746-8. doi: 10.1093/jac/dks111. [PubMed: 22499998].
  • 27. Walsh TJ, Shoham S, Petraitiene R, Sein T, Schaufele R, Kelaher A, et al. Detection of galactomannan antigenemia in patients receiving piperacillin-tazobactam and correlations between in vitro, in vivo, and clinical properties of the drug-antigen interaction. J Clin Microbiol. 2004;42(10):4744-8. doi: 10.1128/JCM.42.10.4744-4748.2004. [PubMed: 15472335].
  • 28. Vergidis P, Razonable RR, Wheat LJ, Estes L, Caliendo AM, Baden LR, et al. Reduction in false-positive Aspergillus serum galactomannan enzyme immunoassay results associated with use of piperacillin-tazobactam in the United States. J Clin Microbiol. 2014;52(6):2199-201. doi: 10.1128/JCM.00285-14. [PubMed: 24719434].
  • 29. Lin JC, Xing YL, Xu WM, Li M, Bo P, Niu YY, et al. Evaluation of galactomannan enzyme immunoassay and quantitative real-time PCR for the diagnosis of invasive pulmonary aspergillosis in a rat model. J Microbiol Biotechnol. 2014;24(8):1044-50. doi: 10.4014/jmb.1401.01067. [PubMed: 24743569].
  • 30. Cuenca-Estrella M, Meije Y, Diaz-Pedroche C, Gomez-Lopez A, Buitrago MJ, Bernal-Martinez L, et al. Value of serial quantification of fungal DNA by a real-time PCR-based technique for early diagnosis of invasive Aspergillosis in patients with febrile neutropenia. J Clin Microbiol. 2009;47(2):379-84. doi: 10.1128/JCM.01716-08. [PubMed: 19109479].
  • 31. Khot PD, Fredricks DN. PCR-based diagnosis of human fungal infections. Expert Rev Anti Infect Ther. 2009;7(10):1201-21. doi: 10.1586/eri.09.104. [PubMed: 19968513].
  • 32. Watzinger F, Suda M, Preuner S, Baumgartinger R, Ebner K, Baskova L, et al. Real-time quantitative PCR assays for detection and monitoring of pathogenic human viruses in immunosuppressed pediatric patients. J Clin Microbiol. 2004;42(11):5189-98. doi: 10.1128/JCM.42.11.5189-5198.2004. [PubMed: 15528714].
  • 33. Nguyen MH, Jaber R, Leather HL, Wingard JR, Staley B, Wheat LJ, et al. Use of bronchoalveolar lavage to detect galactomannan for diagnosis of pulmonary aspergillosis among nonimmunocompromised hosts. J Clin Microbiol. 2007;45(9):2787-92. doi: 10.1128/JCM.00716-07. [PubMed: 17596367].
  • 34. D'Haese J, Theunissen K, Vermeulen E, Schoemans H, De Vlieger G, Lammertijn L, et al. Detection of galactomannan in bronchoalveolar lavage fluid samples of patients at risk for invasive pulmonary aspergillosis: analytical and clinical validity. J Clin Microbiol. 2012;50(4):1258-63. doi: 10.1128/JCM.06423-11. [PubMed: 22301025].
  • 35. Badiee P, Alborzi A, Moeini M, Haddadi P, Farshad S, Japoni A, et al. Antifungal susceptibility of the aspergillus species by Etest and CLSI reference methods. Arch Iran Med. 2012;15(7):429-32. [PubMed: 22724880].
  • 36. Hedayati MT, Mayahi S, Denning DW. A study on Aspergillus species in houses of asthmatic patients from Sari City, Iran and a brief review of the health effects of exposure to indoor Aspergillus. Environ Monit Assess. 2010;168(1-4):481-7. doi: 10.1007/s10661-009-1128-x. [PubMed: 19697147].
  • 37. Zarrinfar H, Mirhendi H, Makimura K, Satoh K, Khodadadi H, Paknejad O. Use of mycological, nested PCR, and real-time PCR methods on BAL fluids for detection of Aspergillus fumigatus and A. flavus in solid organ transplant recipients. Mycopathologia. 2013;176(5-6):377-85. doi: 10.1007/s11046-013-9657-9. [PubMed: 24045934].
  • 38. Zarrinfar H, Mirhendi H, Fata A, Khodadadi H, Kordbacheh P. Detection of Aspergillus flavus and A. fumigatus in Bronchoalveolar Lavage Specimens of Hematopoietic Stem Cell Transplants and Hematological Malignancies Patients by Real-Time Polymerase Chain Reaction, Nested PCR and Mycological Assays. Jundishapur J Microbiol. 2015;8(1). e13744. doi: 10.5812/jjm.13744. [PubMed: 25763133].

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