Is the extraction by Whatman FTA filter matrix technology and sequencing of large ribosomal subunit D1-D2 region sufficient for identification of clinical fungi?

Kiraz N., ÖZ Y., Aslan H., Erturan Z., ENER B., ARIKAN AKDAĞLI S., ...Daha Fazla

MYCOSES, cilt.58, sa.10, ss.588-597, 2015 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 58 Sayı: 10
  • Basım Tarihi: 2015
  • Doi Numarası: 10.1111/myc.12365
  • Dergi Adı: MYCOSES
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.588-597
  • Anahtar Kelimeler: Aspergillus, Candida, D1-D2 region, Fusarium, identification, Whatman FTA filter, DESORPTION IONIZATION-TIME, MEDICALLY IMPORTANT YEASTS, FLIGHT MASS-SPECTROMETRY, TRANSCRIBED SPACER REGIONS, CANDIDA-ALBICANS, MOLECULAR-IDENTIFICATION, RAPID IDENTIFICATION, DNA EXTRACTION, PCR, FUSARIUM
  • Eskişehir Osmangazi Üniversitesi Adresli: Evet

Özet

Although conventional identification of pathogenic fungi is based on the combination of tests evaluating their morphological and biochemical characteristics, they can fail to identify the less common species or the differentiation of closely related species. In addition these tests are time consuming, labour-intensive and require experienced personnel. We evaluated the feasibility and sufficiency of DNA extraction by Whatman FTA filter matrix technology and DNA sequencing of D1-D2 region of the large ribosomal subunit gene for identification of clinical isolates of 21 yeast and 160 moulds in our clinical mycology laboratory. While the yeast isolates were identified at species level with 100% homology, 102 (63.75%) clinically important mould isolates were identified at species level, 56 (35%) isolates at genus level against fungal sequences existing in DNA databases and two (1.25%) isolates could not be identified. Consequently, Whatman FTA filter matrix technology was a useful method for extraction of fungal DNA; extremely rapid, practical and successful. Sequence analysis strategy of D1-D2 region of the large ribosomal subunit gene was found considerably sufficient in identification to genus level for the most clinical fungi. However, the identification to species level and especially discrimination of closely related species may require additional analysis.