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2016 ; 54
(11
): 2798-2803
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gab.com Text
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Discrimination of Aspergillosis, Mucormycosis, Fusariosis, and Scedosporiosis in
Formalin-Fixed Paraffin-Embedded Tissue Specimens by Use of Multiple Real-Time
Quantitative PCR Assays
#MMPMID27605714
Salehi E
; Hedayati MT
; Zoll J
; Rafati H
; Ghasemi M
; Doroudinia A
; Abastabar M
; Tolooe A
; Snelders E
; van der Lee HA
; Rijs AJ
; Verweij PE
; Seyedmousavi S
; Melchers WJ
J Clin Microbiol
2016[Nov]; 54
(11
): 2798-2803
PMID27605714
show ga
In a retrospective multicenter study, 102 formalin-fixed paraffin-embedded (FFPE)
tissue specimens with histopathology results were tested. Two 4- to 5-?m FFPE
tissue sections from each specimen were digested with proteinase K, followed by
automated nucleic acid extraction. Multiple real-time quantitative PCR (qPCR)
assays targeting the internal transcribed spacer 2 (ITS2) region of ribosomal
DNA, using fluorescently labeled primers, was performed to identify clinically
important genera and species of Aspergillus, Fusarium, Scedosporium, and the
Mucormycetes The molecular identification was correlated with results from
histological examination. One of the main findings of our study was the high
sensitivity of the automated DNA extraction method, which was estimated to be
94%. The qPCR procedure that was evaluated identified a range of fungal
genera/species, including Aspergillus fumigatus, Aspergillus flavus, Aspergillus
terreus, Aspergillus niger, Fusarium oxysporum, Fusarium solani, Scedosporium
apiospermum, Rhizopus oryzae, Rhizopus microsporus, Mucor spp., and
Syncephalastrum Fusarium oxysporum and F. solani DNA was amplified from five
specimens from patients initially diagnosed by histopathology as having
aspergillosis. Aspergillus flavus, S. apiospermum, and Syncephalastrum were
detected from histopathological mucormycosis samples. In addition, examination of
four samples from patients suspected of having concomitant aspergillosis and
mucormycosis infections resulted in the identification of two A. flavus isolates,
one Mucor isolate, and only one sample having both R. oryzae and A. flavus Our
results indicate that histopathological features of molds may be easily confused
in tissue sections. The qPCR assay used in this study is a reliable tool for the
rapid and accurate identification of fungal pathogens to the genus and species
levels directly from FFPE tissues.