Hostname: page-component-cd9895bd7-mkpzs Total loading time: 0 Render date: 2024-12-28T00:59:51.208Z Has data issue: false hasContentIssue false
  • English
  • Français

Identification of pathogens causing invasive fungal rhinosinusitis in surgical biopsies using polymerase chain reaction

Published online by Cambridge University Press:  20 July 2020

S Chaturantabut ,
N Kitkumthorn ,
A Mutirangura ,
N Praditphol ,
A Chindamporn ,
P S Thorner  and
S Keelawat
S Chaturantabut
Affiliation:
Medical Science Program, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
N Kitkumthorn
Affiliation:
Department of Oral Biology, Faculty of Dentistry, Mahidol University, Bangkok, Thailand
A Mutirangura
Affiliation:
Department of Anatomy, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
N Praditphol
Affiliation:
Department of Pathology, Rajavithi Hospital, Bangkok, Thailand
A Chindamporn
Affiliation:
Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
P S Thorner
Affiliation:
Department of Laboratory Medicine and Pathobiology, University of Toronto, Canada Department of Pathology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
S Keelawat*
Affiliation:
Department of Pathology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
*
Author for correspondence: Dr Somboon Keelawat, Department of Pathology, Faculty of Medicine, Chulalongkorn University, 1873 King Rama IV Street, Bangkok, 10330Thailand E-mail: trcskl@gmail.com
Get access Rights & Permissions [Opens in a new window]

Abstract

Background

Invasive fungal rhinosinusitis is associated with high morbidity and mortality. Rapid pathogen identification is mandatory, but fresh tissue is not always available. A polymerase chain reaction method was designed in order to detect fungi in formalin-fixed paraffin-embedded samples. This was applied to a retrospective series of tissue biopsies from Thai patients with invasive fungal rhinosinusitis.

Methods

Tissue blocks from 64 cases yielded adequate DNA. Three sequential polymerase chain reaction were performed: ZP3 (housekeeping gene) and panfungal polymerase chain reactions, and a differentiating polymerase chain reaction based on the 5.8s ribosomal RNA and internal transcribed spacer 2 regions. The polymerase chain reaction products were then sequenced.

Results

Polymerase chain reaction identified a fungal pathogen in 20 of 64 cases (31 per cent). Aspergillus species was the most common cause of invasive fungal rhinosinusitis (nine cases). Other causes included candida (n = 4), cladosporium (n = 4), mucor (n = 1), alternaria (n = 1) and dendryphiella (n = 1) species.

Conclusion

Polymerase chain reaction can provide rapid identification of fungal pathogens in paraffin-embedded tissue, enabling prompt treatment of invasive fungal rhinosinusitis.

Keywords

Invasive Fungal RhinosinusitisPolymerase Chain ReactionAspergillusCandidaCladosporium
Type
Main Articles
Information
The Journal of Laryngology & Otology , Volume 134 , Issue 7 , July 2020 , pp. 632 - 635
Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

Dr S Keelawat takes responsibility for the integrity of the content of the paper

References

Das, A, Bal, A, Chakrabarti, A, Panda, N, Joshi, K. Spectrum of fungal rhinosinusitis; histopathologist's perspective. Histopathology 2009;54:854–9CrossRefGoogle ScholarPubMed
Taxy, JB. Paranasal fungal sinusitis: contributions of histopathology to diagnosis: a report of 60 cases and literature review. Am J Surg Pathol 2006;30:713–20CrossRefGoogle ScholarPubMed
Chen, CY, Sheng, WH, Cheng, A, Chen, YC, Tsay, W, Tang, JL et al. Invasive fungal sinusitis in patients with hematological malignancy: 15 years experience in a single university hospital in Taiwan. BMC Infect Dis 2011;11:250CrossRefGoogle Scholar
Iwen, PC, Rupp, ME, Hinrichs, SH. Invasive mold sinusitis: 17 cases in immunocompromised patients and review of the literature. Clin Infect Dis 1997;24:1178–84CrossRefGoogle ScholarPubMed
Schell, WA. Histopathology of fungal rhinosinusitis. Otolaryngol Clin North Am 2000;33:251–76CrossRefGoogle ScholarPubMed
Gillespie, MB, O'Malley, BW. An algorithmic approach to the diagnosis and management of invasive fungal rhinosinusitis in the immunocompromised patient. Otolaryngol Clin North Am 2000;33:323–34CrossRefGoogle ScholarPubMed
Groppo, ER, El-Sayed, IH, Aiken, AH, Glastonbury, CM. Computed tomography and magnetic resonance imaging characteristics of acute invasive fungal sinusitis. Arch Otolaryngol Head Neck Surg 2011;137:1005–10CrossRefGoogle ScholarPubMed
Lai, CC, Tan, CK, Huang, YT, Shao, PL, Hsueh, PR. Current challenges in the management of invasive fungal infections. J Infect Chemother 2008;14:77–85CrossRefGoogle ScholarPubMed
Stergiopoulou, T, Walsh, TJ. Clinical pharmacology of antifungal agents to overcome drug resistance in pediatric patients. Expert Opin Pharmacother 2015;16:213–26CrossRefGoogle ScholarPubMed
Ruangritchankul, K, Chindamporn, A, Worasilchai, N, Poumsuk, U, Keelawat, S, Bychkov, A. Invasive fungal disease in university hospital: a PCR-based study of autopsy cases. Int J Clin Exp Pathol 2015;8:14840–52Google ScholarPubMed
Babouee Flury, B, Weisser, M, Prince, SS, Bubendorf, L, Battegay, M, Frei, R et al. Performances of two different panfungal PCRs to detect mould DNA in formalin-fixed paraffin-embedded tissue: what are the limiting factors? BMC Infect Dis 2014;14:692–700CrossRefGoogle ScholarPubMed
Chamilos, G, Luna, M, Lewis, RE, Bodey, GP, Chemaly, R, Tarrand, JJ et al. Invasive fungal infections in patients with hematologic malignancies in a tertiary care cancer center: an autopsy study over a 15-year period (1989–2003). Haematologica 2006;91:986–9Google Scholar
Rickerts, V, Mousset, S, Lambrecht, E, Tintelnot, K, Schwerdtfeger, R, Presterl, E et al. Comparison of histopathological analysis, culture, and polymerase chain reaction assays to detect invasive mold infections from biopsy specimens. Clin Infect Dis 2007;44:1078–83CrossRefGoogle ScholarPubMed
Alexander, BD. Diagnosis of fungal infection: new technologies for the mycology laboratory. Transpl Infect Dis 2002;4(suppl 3):32–7CrossRefGoogle ScholarPubMed
Avni, T, Leibovici, L, Paul, M. PCR diagnosis of invasive candidiasis: systematic review and meta-analysis. J Clin Microbiol 2011;49:665–70CrossRefGoogle ScholarPubMed
Baskova, L, Buchta, V. Laboratory diagnostics of invasive fungal infections: an overview with emphasis on molecular approach. Folia Microbiol (Praha) 2012;57:421–30CrossRefGoogle ScholarPubMed
Bialek, R, Konrad, F, Kern, J, Aepinus, C, Cecenas, L, Gonzalez, GM et al. PCR based identification and discrimination of agents of mucormycosis and aspergillosis in paraffin wax embedded tissue. J Clin Pathol 2005;58:1180–4CrossRefGoogle ScholarPubMed
Borman, AM, Linton, CJ, Miles, SJ, Johnson, EM. Molecular identification of pathogenic fungi. J Antimicrob Chemother 2008;61(suppl 1):i7–12CrossRefGoogle ScholarPubMed
Buitrago, MJ, Bernal-Martinez, L, Castelli, MV, Rodriguez-Tudela, JL, Cuenca-Estrella, M. Performance of panfungal - and specific - PCR-based procedures for etiological diagnosis of invasive fungal diseases on tissue biopsy specimens with proven infection: a 7-year retrospective analysis from a reference laboratory. J Clin Microbiol 2014;52:1737–40CrossRefGoogle ScholarPubMed
Gharizadeh, B, Norberg, E, Loffler, J, Jalal, S, Tollemar, J, Einsele, H et al. Identification of medically important fungi by the pyrosequencing technology. Mycoses 2004;47:29–33CrossRefGoogle ScholarPubMed
Gholinejad-Ghadi, N, Shokohi, T, Seifi, Z, Aghili, SR, Roilides, E, Nikkhah, M et al. Identification of mucorales in patients with proven invasive mucormycosis by polymerase chain reaction in tissue samples. Mycoses 2018;61:909–15CrossRefGoogle ScholarPubMed
Khot, PD, Fredricks, DN. PCR-based diagnosis of human fungal infections. Expert Rev Anti Infect Ther 2009;7:1201–21CrossRefGoogle ScholarPubMed
Kourkoumpetis, TK, Fuchs, BB, Coleman, JJ, Desalermos, A, Mylonakis, E. Polymerase chain reaction-based assays for the diagnosis of invasive fungal infections. Clin Infect Dis 2012;54:1322–31CrossRefGoogle Scholar
Munoz-Cadavid, C, Rudd, S, Zaki, SR, Patel, M, Moser, SA, Brandt, ME et al. Improving molecular detection of fungal DNA in formalin-fixed paraffin-embedded tissues: comparison of five tissue DNA extraction methods using panfungal PCR. J Clin Microbiol 2010;48:2147–53CrossRefGoogle ScholarPubMed
Salehi, E, Hedayati, MT, Zoll, J, Rafati, H, Ghasemi, M, Doroudinia, A et al. Discrimination of aspergillosis, mucormycosis, fusariosis, and scedosporiosis in formalin-fixed paraffin-embedded tissue specimens by use of multiple real-time quantitative PCR assays. J Clin Microbiol 2016;54:2798–803CrossRefGoogle ScholarPubMed
Sambrook, J, Russell, DW. Purification of nucleic acids by extraction with phenol:chloroform. CSH Protoc 2006;2006:pdb.prot4455Google ScholarPubMed
Praweswararat, P. Invasive fungal sinusitis: seven years experiences in Rajavithi Hospital. Asian Rhinology Journal 2015;2:33–9Google Scholar
Roongrotwattanasiri, K, Fooanant, S, Naksane, L. Invasive fungal rhinosinusitis in Maharaj Nakorn Chiang Mai Hospital. J Med Assoc Thai 2007;90:2524–8Google ScholarPubMed
Soontrapa, P, Larbcharoensub, N, Luxameechanporn, T, Cheewaruangroj, W, Prakunhungsit, S, Sathapatayavong, B et al. Fungal rhinosinusitis: a retrospective analysis of clinicopathologic features and treatment outcomes at Ramathibodi Hospital. Southeast Asian J Trop Med Public Health 2010;41:442–9Google ScholarPubMed
Michael, RC, Michael, JS, Ashbee, RH, Mathews, MS. Mycological profile of fungal sinusitis: an audit of specimens over a 7-year period in a tertiary care hospital in Tamil Nadu. Indian J Pathol Microbiol 2008;51:493–6CrossRefGoogle Scholar
Bhatt, VR, Viola, GM, Ferrajoli, A. Invasive fungal infections in acute leukemia. Ther Adv Hematol 2011;2:231–47CrossRefGoogle ScholarPubMed
Pastor, FJ, Guarro, J. Alternaria infections: laboratory diagnosis and relevant clinical features. Clin Microbiol Infect 2008;14:734–46CrossRefGoogle ScholarPubMed
Hamilos, DL. Allergic fungal rhinitis and rhinosinusitis. Proc Am Thorac Soc 2010;7:245–52CrossRefGoogle ScholarPubMed
Lee, B, Baker, G. Environment and the distribution of microfungi in a Hawaiian mangrove swamp. Pacific Science 1972;26:11–19Google Scholar
Lau, A, Chen, S, Sorrell, T, Carter, D, Malik, R, Martin, P et al. Development and clinical application of a panfungal PCR assay to detect and identify fungal DNA in tissue specimens. J Clin Microbiol 2007;45:380–5CrossRefGoogle ScholarPubMed