title: Comparison between the effectiveness of polymerase chain reaction and in situ hybridization in detecting the presence of pathogenic fungi by using the preserved DNA in formalin-fixed and paraffin-embedded tissues.
abstract: In situ hybridization (ISH) has been recognized as an important technique for identifying the causative fungi in the foci of infection observed in histopathological specimens which was processed from formalin-fixed and paraffin-embedded (FFPE) tissues. However, few basic studies have conducted an evaluation of the DNA preservation for use in ISH in comparison to polymerase chain reaction (PCR). The latter is a DNA amplification-based modality. In the present study, we analyzed 65 FFPE lung tissue specimens collected from autopsy cases for comparing the usefulness of ISH and PCR analysis. As a result, the positive identification rates for PCR were strikingly low; a majority of these results can be assumed to be false negative because the presence of fungi had been confirmed by histopathological analysis. In contrast, panfungal ISH targeting of the 28S rRNA showed a higher sensitivity than the 230-bp panfungal PCR primers did (80.0% versus 4.6%, respectively). Furthermore, over 60% of the samples we examined showed a favorable intensity of the ISH signal. Therefore, in conventional postmortem FFPE tissues, the state of DNA preservation may be more favorable for ISH than PCR analysis.
authors.affiliation: Department of Surgical Pathology, Toho University School of Medicine, Tokyo, Japan.
authors.forename: Yuzhu
authors.initials: Y
authors.lastname: Zhi
authors.forename: Daisuke
authors.initials: D
authors.lastname: Sasai
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Okubo
authors.forename: Minoru
authors.initials: M
authors.lastname: Shinozaki
authors.forename: Haruo
authors.initials: H
authors.lastname: Nakayama
authors.forename: Somay
authors.initials: S
authors.lastname: Yamagata Murayama
authors.forename: Megumi
authors.initials: M
authors.lastname: Wakayama
authors.forename: Tadashi
authors.initials: T
authors.lastname: Ide
authors.forename: Zean
authors.initials: Z
authors.lastname: Zhang
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Shibuya
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D004271:DNA, Fungal / Q000235:genetics; D005260:Female; D005658:Fungi / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D007223:Infant; D008297:Male; D008875:Middle Aged; D009172:Mycology / Q000379:methods*; D009181:Mycoses / Q000175:diagnosis* / Q000382:microbiology; D057089:Pathology, Molecular / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D004271:DNA, Fungal