Identification of Pseudallescheria and Scedosporium species by three molecular methods

Lu, Q. and Gerrits Van Den Ende, A. H. G. and Bakkers, J. M. J. E. and Sun, J. and Lackner, M. and Najafzadeh, M. J. and Melchers, W. J. G. and Li, R. and De Hoog, G. S. (2011) Identification of Pseudallescheria and Scedosporium species by three molecular methods. Journal of Clinical Microbiology, 49 (3). pp. 960-967.

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Abstract

The major clinically relevant species in Scedosporium (teleomorph Pseudallescheria) are Pseudallescheria boydii, Scedosporium aurantiacum, Scedosporium apiospermum, and Scedosporium prolificans, while Pseudallescheria minutispora, Petriellopsis desertorum, and Scedosporium dehoogii are exceptional agents of disease. Three molecular methods targeting the partial β-tubulin gene were developed and evaluated to identify six closely related species of the S. apiospermum complex using quantitative real-time PCR (qPCR), PCR-based reverse line blot (PCR-RLB), and loop-mediated isothermal amplification (LAMP). qPCR was not specific enough for the identification of all species but had the highest sensitivity. The PCR-RLB assay was efficient for the identification of five species. LAMP distinguished all six species unambiguously. The analytical sensitivities of qPCR, PCR-RLB, and LAMP combined with MagNAPure, CTAB (cetyltrimethylammonium bromide), and FTA filter (Whatman) extraction were 50, 5 × 103, and 5 × 102 cells/μl, respectively. When LAMP was combined with a simplified DNA extraction method using an FTA filter, identification to the species level was achieved within 2 h, including DNA extraction. The FTA-LAMP assay is therefore recommended as a cost-effective, simple, and rapid method for the identification of Scedosporium species. Copyright © 2011, American Society for Microbiology. All Rights Reserved.

Item Type: Article
Additional Information: Cited By :39 Export Date: 16 February 2020 CODEN: JCMID Correspondence Address: Li, R.; Department of Dermatology, Peking University First Hospital, Peking University, No. 8 Xishiku St., West District, 100034 Beijing, China; email: lry0660@gmail.com
Uncontrolled Keywords: beta tubulin cetrimide article Ascomycetes controlled study cost effectiveness analysis DNA extraction fungus identification gene amplification loop mediated isothermal amplification nonhuman nucleotide sequence PCR based reverse line blot Petriellidium boydii Petriellopsis desertorum priority journal Pseudallescheria minutispora real time polymerase chain reaction Scedosporium Scedosporium apiospermum Scedosporium aurantiacum Scedosporium dehoogii Scedosporium prolificans sensitivity and specificity species identification teleomorph Fungal Proteins Humans Molecular Diagnostic Techniques Mycetoma Mycology Nucleic Acid Amplification Techniques Pseudallescheria Tubulin Pseudallescheria boydii
Subjects: QW Microbiology and Immunology
Divisions: Mashhad University of Medical Sciences
Depositing User: mr lib7 lib7
Date Deposited: 29 Apr 2020 05:09
Last Modified: 29 Apr 2020 05:09
URI: http://eprints.mums.ac.ir/id/eprint/17044

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