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Emergence of Aureobasidium pullulans as human fungal pathogen and molecular assay for future medical diagnosis

Folia Microbiologica volume 56, Article number: 459 (2011) Cite this article

Abstract

Despite the great importance of Aureobasidium pullulans in biotechnology, the fungus had emerged as an opportunistic human pathogen, especially among immunocompromised patients. Clinical detection of this rare human fungal pathogen presently relies on morphology diagnosis which may be misleading. Thus, a sensitive and accurate quantitative molecular assay for A. pullulans remains lacking. In this study, we presented the microscopy observations of A. pullulans that reveals the phenotypic plasticity of the fungus. A. pullulans-specific primers and molecular beacon probes were designed based on the fungal 18S ribosomal RNA (rRNA) gene. Comparison of two probes with varied quencher chemistry, namely BHQ-1 and Tamra, revealed high amplification efficiency of 104% and 108%, respectively. The optimized quantitative real-time PCR (qPCR) assays could detect and quantify up to 1 pg concentration of A. pullulans DNA. Both assays displayed satisfactory performance parameters at fast thermal cycling mode. The molecular assay has great potential as a molecular diagnosis tool for early detection of fungal infection caused by A. pullulans, which merits future study in clinical diagnosis.

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References

  • Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402. doi:10.1093/nar/25.17.3389

    PubMed  Article  CAS  Google Scholar 

  • Barnes RA (2008) Early diagnosis of fungal infection in immunocompromised patients. J Antimicrob Chemoth 61(Suppl 1):i3–i6. doi:10.1093/jac/dkm424

    Article  CAS  Google Scholar 

  • Bolignano G, Criseo G (2003) Disseminated nosocomial fungal infection by Aureobasidium pullulans var. melanigenum: a case report. J Clin Microbiol 41:4483–4485. doi:10.1128/JCM.41.9.4483-4485.2003

    PubMed  Article  Google Scholar 

  • Chan GF, Puad MSA, Rashid NAA (2011) Candida orthopsilosis and Aureobasidium pullulans—rare fungal pathogens causing persistent skin infection. Insight Infectious Diseases 1:1–4. doi:10.5567/INFECTIOUS-IK.2011.1.4

    Article  Google Scholar 

  • Chi ZM, Wang F, Chi Z, Yue LX, Liu GL, Zhang T (2009) Bioproducts from Aureobasidium pullulans, a biotechnologically important yeast. Appl Microbiol Biotechnol 82:793–804. doi:10.1007/s00253-009-1882-2

    PubMed  Article  CAS  Google Scholar 

  • Clark EC, Silver SM, Hollick GE, Rinaldi MG (1995) Continuous ambulatory peritoneal dialysis complicated by Aureobasidium pullulans peritonitis. Am J Nephrol 15:353–355. doi:10.1159/000168863

    PubMed  Article  CAS  Google Scholar 

  • Cooke WB (1959) An ecological life history of Aureobasidium pullulans (de Bary) Arnaud. Mycopathologia 12:1–45

    PubMed  CAS  Google Scholar 

  • Guarner J, Brandt ME (2011) Histopathologic diagnosis of fungal infections in the 21st century. Clin Microbiol Rev 24:247–280. doi:10.1128/CMR.00053-10

    PubMed  Article  Google Scholar 

  • Hawkes M, Rennie R, Sand C, Vaudry W (2005) Aureobasidium pullulans infection: fungemia in an infant and a review of human cases. Diagn Micr Infec Dis 51:209–213. doi:10.1016/j.diagmicrobio.2004.10.007

    Article  Google Scholar 

  • Huang Y-T, Liao C-H, Huang Y-T, Liaw S-J, Yang J-L, Lai D-M, Lee Y-C, Hsueh P-R (2008) Catheter-related septicemia due to Aureobasidium pullulans. Int J Infec Dis 12:e137–e139. doi:10.1016/j.ijid.2008.02.004

    Article  Google Scholar 

  • Jones FR, Christensen GR (1974) Pullularia corneal ulcer. Arch Ophthalmol 92:529–530

    PubMed  CAS  Google Scholar 

  • Joshi A, Singh R, Shah MS, Umesh S, Khattry N (2010) Subcutaneous mycosis and fungemia by Aureobasidium pullulans: a rare pathogenic fungus in a post allogeneic BM transplant patient. Bone Marrow Transpl 45:203–204. doi:10.1038/bmt.2009.111

    Article  CAS  Google Scholar 

  • La Paz JL, Esteve T, Pla M (2007) Comparison of real-time PCR detection chemistries and cycling modes using Mon810 event-specific assays as model. J Agric Food Chem 55:4312–4318. doi:10.1021/jf063725g

    PubMed  Article  Google Scholar 

  • Li S, Cullen D, Hjort M, Spear R, Andrews JH (1996) Development of an oligonucleotide probe for Aureobasidium pullulans based on the small-subunit rRNA gene. Appl Environ Microbiol 62:1514–1518

    PubMed  CAS  Google Scholar 

  • Manitchotpisit P, Price NPJ, Leathers TD, Punnapayak H (2011) Heavy oils produced by Aureobasidium pullulans. Biotechnol Lett 33:1151–1157. doi:10.1007/s10529-011-0548-1

    PubMed  Article  CAS  Google Scholar 

  • Markham NR, Zuker M (2005) DINAMelt web server for nucleic acid melting prediction. Nucleic Acids Res 33:W577–W581. doi:10.1093/nar/gki591

    PubMed  Article  CAS  Google Scholar 

  • Niedoszytko M, Chelminska M, Jassem E, Czestochowska E (2007) Association between sensitization to Aureobasidium pullulans (Pullularia sp) and severity of asthma. Ann Allergy Asthma Immunol 98:153–156

    PubMed  Article  Google Scholar 

  • Panda A, Das H, Deb M, Khanal B, Kumar S (2006) Aureobasidium pullulans keratitis. Clin Exp Ophthalmol 34:260–264. doi:10.1111/j.1442-9071.2006.1201.x

    Article  Google Scholar 

  • Pikazis D, Xynos ID, Xila V, Velegraki A, Aroni K (2009) Extended fungal skin infection due to Aureobasidium pullulans. Clin Exp Dermatol 34:e892–e894. doi:10.1111/j.1365-2230.2009.03663.x

    PubMed  Article  CAS  Google Scholar 

  • Prasongsuk S, Sullivan RF, Kuhirun M, Eveleigh DE, Punnapayak H (2005) Thailand habitats as sources of pullulan-producing strains of Aureobasidium pullulans. World J Microb Biot 21:393–398. doi:10.1007/s11274-004-2237-x

    Article  CAS  Google Scholar 

  • Pritchard RC, Muir DB (1987) Black fungi: a survey of dematiaceous hyphomycetes from clinical specimens identified over a five year period in a reference laboratory. Pathology 19:281–284. doi:10.3109/00313028709066564

    PubMed  Article  CAS  Google Scholar 

  • Ravella SA, Quiñones TS, Retter A, Heiermann M, Amon T, Hobbs PJ (2010) Extracellular polysaccharide (EPS) production by a novel strain of yeast-like fungus Aureobasidium pullulans. Carbohyd Polym 82:728–732. doi:10.1016/j.carbpol.2010.05.039

    Article  CAS  Google Scholar 

  • Redondo-Bellόn P, Idoate M, Rubio M, Ignacio Herrero J (1997) Chromoblastomycosis produced by Aureobasidium pullulans in an immunosuppressed patient. Arch Dermatol 133:663–664

    Article  Google Scholar 

  • Rozen S, Skaletsky HJ (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols: methods in molecular biology. Humana, Totowa, pp 365–386

    Google Scholar 

  • Rumbold K, Biely P, Mastihubová M, Gudelj M, Gübitz G, Robra K-H, Prior BA (2003) Purification and properties of a feruloyl esterase involved in lignocellulose degradation by Aureobasidium pullulan. Appl Environ Microbiol 69:5622–5626. doi:10.1128/AEM.69.9.5622-5626.2003

    PubMed  Article  CAS  Google Scholar 

  • Salkin IF, Martinez JA, Kemna ME (1986) Opportunistic infection of the spleen caused by Aureobasidium pullulans. J Clin Microbiol 23:828–831

    PubMed  CAS  Google Scholar 

  • Schena L, Sialer MF, Gallitelli D (2002) Molecular detection of strain L47 of Aureobasidium pullulans, a biocontrol agent of postharvest diseases. Plant Disease 86:54–60

    Article  CAS  Google Scholar 

  • Schmid F, Moser G, Muller H, Berg G (2011) Functional and structural microbial diversity in organic and conventional viticulture: organic farming benefits natural biocontrol agents. Appl Environ Microbiol 77:2188–2191. doi:10.1128/AEM.02187-10

    PubMed  Article  CAS  Google Scholar 

  • Slepecky RA, Starmer WT (2009) Phenotypic plasticity in fungi: a review with observations on Aureobasidium pullulans. Mycologia 101:823–832. doi:10.3852/08-197

    PubMed  Article  Google Scholar 

  • Taylor PE, Esch R, Flagan RC, House J, Tran L, Glovsky MM (2006) Identification and possible disease mechanisms of an under-recognized fungus, Aureobasidium pullulans. Int Arch Allergy Immunol 139:45–52. doi:10.1159/000089522

    PubMed  Article  Google Scholar 

  • Vollmer T, Stormer M, Kleesiek K, Dreier J (2008) Evaluation of novel broad-range real-time PCR assay for rapid detection of human pathogenic fungi in various clinical specimens. J Clin Microbiol 46:1919–1926. doi:10.1128/JCM.02178-07

    PubMed  Article  CAS  Google Scholar 

  • Wilson SJ, Everts RJ, Kirkland KB, Sexton DJ (2000) A pseudo-outbreak of Aureobasidium species lower respiratory tract infections caused by reuse of single-use stopcocks during bronchoscopy. Infect Control Hos Epidemiol 21:470–472

    Article  CAS  Google Scholar 

  • Wu Z, Tsumura Y, Blomquist G, Wang X-R (2003) 18S rRNA gene variation among common airborne fungi, and development of specific oligonucleotide probes for the detection of fungal isolates. Appl Environ Microbiol 69:5389–5397. doi:10.1128/AEM.69.9.5389-5397.2003

    PubMed  Article  CAS  Google Scholar 

  • Zalar P, Gostinčar C, de Hoog GS, Uršič V, Sudhadham M, Gunde-Cimerman N (2008) Redefinition of Aureobasidium pullulans and its varieties. Stud Mycol 61:21–38. doi:10.3114/sim.2008.61.02

    PubMed  Article  CAS  Google Scholar 

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Acknowledgments

This research was supported by the Research University Grant Scheme of Universiti Teknologi Malaysia.

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Affiliations

  1. Nanoporous Materials for Biological Application Research Group (NAMBAR), Sustainability Research Alliance, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia

    Giek Far Chan, Mohamad Safwan Ahmad Puad, Chai Fung Chin & Noor Aini Abdul Rashid

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  1. Giek Far Chan
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  2. Mohamad Safwan Ahmad Puad
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  3. Chai Fung Chin
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  4. Noor Aini Abdul Rashid
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Correspondence to Giek Far Chan.

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Chan, G.F., Puad, M.S.A., Chin, C.F. et al. Emergence of Aureobasidium pullulans as human fungal pathogen and molecular assay for future medical diagnosis. Folia Microbiol 56, 459 (2011). https://doi.org/10.1007/s12223-011-0070-9

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  • DOI: https://doi.org/10.1007/s12223-011-0070-9

Keywords

  • Invasive Fungal Infection
  • Molecular Beacon
  • Invasive Fungal Disease
  • Sabouraud Dextrose Agar
  • Aureobasidium Pullulans