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Abstract

Purpose. Tissue samples from patients with suspicion of deep or subcutaneous fungal infections were analysed at the Portuguese Reference Mycology Laboratory according to a proposed diagnostic approach, which aims to constitute a rapid and accurate diagnosis for these fungal infections.

Methodology. Forty-six tissue biopsy samples were analysed over a period of 26 months, using a diagnostic approach that includes culture, panfungal PCR and Aspergillus-directed PCR.

Results/Key findings. Overall, 23 samples were reported as negative while the remaining 23 were reported as positive for fungi (PCR, culture and/or histology). PCR showed an estimated detection limit of 12 pg DNA µl. From the 46 samples, 30 were negative for fungal DNA while 16 gave positive results. From these, 12 cases were detected by panfungal PCR and six cases by PCR directed toward Aspergillus. In 61 % of the cases, there was concordance between molecular and cultural methods. Aetiological agents identified were Candida albicans, C. glabrata, C. tropicalis, Trichosporon montevideense, Alternaria spp., Exophiala sp., Trichoderma sp., Histoplasma spp., Aspergillus fumigatus, Trichophyton rubrum and Paracoccidioides brasiliensis.

Conclusion. Our results showed that the proposed polyphasic approach appears to be a useful strategy in the detection of fungi from tissue samples, allowing a better prognosis. In further studies, the inclusion of a higher number of samples and the implementation of more genus-specific PCRs will certainly contribute to an increase in the specificity and sensitivity of this method.

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2018-11-27
2019-10-22
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References

  1. Badiee P, Hashemizadeh Z. Opportunistic invasive fungal infections: diagnosis & clinical management. Indian J Med Res 2014;139:195–204[PubMed]
    [Google Scholar]
  2. De Pauw B, Walsh TJ, Donnelly JP, Stevens DA, Edwards JE 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;46:1813–1821 doi [CrossRef][PubMed]
    [Google Scholar]
  3. Clinical and Laboratory Standards Institute (CLSI) Principles and procedures for detection of fungi in clinical specimens- direct examination and culture; Approved guideline. CLSI Document M54-A (Isbn 1-56238-858-4) 96 West Valley Road, Suite 2500, Wayne, Pennsylvania 19087 USA: Clinical and Laboratory Standards Institute; 2012
    [Google Scholar]
  4. Hoog DC, Guarro J, Gene G, Figueras M. Atlas of Clinical Fungi &Ndash; the Ultimate Benchtool for DiagnosisVersion 4.1.4 Utrecht: Centraalbureau voor Schimmelcultures; 2016
    [Google Scholar]
  5. Buitrago MJ, Aguado JM, Ballen A, Bernal-Martinez L, Prieto M et al. Efficacy of DNA amplification in tissue biopsy samples to improve the detection of invasive fungal disease. Clin Microbiol Infect 2013;19:E271E277 [CrossRef][PubMed]
    [Google Scholar]
  6. White TJ, Bruns T, Lee S, Taylor JW. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In Innis MA, Gelfand DH, Sninsky JJ, White TJ. (editors) PCR Protocols: A Guide To Methods And Applications New York: Academic Press, Inc; 1990; pp.315–322
    [Google Scholar]
  7. Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 1994;22:4673–4680 [CrossRef][PubMed]
    [Google Scholar]
  8. Babouee B, Goldenberger D, Elzi L, Lardinois D, Sadowski-Cron C et al. Prospective study of a panfungal PCR assay followed by sequencing, for the detection of fungal DNA in normally sterile specimens in a clinical setting: a complementary tool in the diagnosis of invasive fungal disease?. Clin Microbiol Infect 2013;19:e354e357 [CrossRef][PubMed]
    [Google Scholar]
  9. Ala-Houhala M, Koukila-Kähkölä P, Antikainen J, Valve J, Kirveskari J et al. Clinical use of fungal PCR from deep tissue samples in the diagnosis of invasive fungal diseases: a retrospective observational study. Clin Microbiol Infect 2018;24:301–305 [CrossRef][PubMed]
    [Google Scholar]
  10. Trubiano JA, Dennison AM, Morrissey CO, Chua KY, Halliday CL et al. Clinical utility of panfungal polymerase chain reaction for the diagnosis of invasive fungal disease: a single center experience. Med Mycol 2016;54:138–146 [CrossRef][PubMed]
    [Google Scholar]
  11. Posteraro B, Torelli R, de Carolis E, Posteraro P, Sanguinetti M. Antifungal susceptibility testing: current role from the clinical laboratory perspective. Mediterr J Hematol Infect Dis 2014;6:e2014030 [CrossRef][PubMed]
    [Google Scholar]
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