RRC ID 46774
著者 Muraosa Y, Toyotome T, Yahiro M, Watanabe A, Shikanai-Yasuda MA, Kamei K.
タイトル Detection of Histoplasma capsulatum from clinical specimens by cycling probe-based real-time PCR and nested real-time PCR.
ジャーナル Med Mycol
Abstract We developed new cycling probe-based real-time PCR and nested real-time PCR assays for the detection of Histoplasma capsulatum that were designed to detect the gene encoding N-acetylated α-linked acidic dipeptidase (NAALADase), which we previously identified as an H. capsulatum antigen reacting with sera from patients with histoplasmosis. Both assays specifically detected the DNAs of all H. capsulatum strains but not those of other fungi or human DNA. The limited of detection (LOD) of the real-time PCR assay was 10 DNA copies when using 10-fold serial dilutions of the standard plasmid DNA and 50 DNA copies when using human serum spiked with standard plasmid DNA. The nested real-time PCR improved the LOD to 5 DNA copies when using human serum spiked with standard plasmid DNA, which represents a 10-fold higher than that observed with the real-time PCR assay. To assess the ability of the two assays to diagnose histoplasmosis, we analyzed a small number of clinical specimens collected from five patients with histoplasmosis, such as sera (n = 4), formalin-fixed paraffin-embedded (FFPE) tissue (n = 4), and bronchoalveolar lavage fluid (BALF) (n = 1). Although clinical sensitivity of the real-time PCR assay was insufficiently sensitive (33%), the nested real-time PCR assay increased the clinical sensitivity (77%), suggesting it has a potential to be a useful method for detecting H. capsulatum DNA in clinical specimens.
巻・号 54(4)
ページ 433-8
公開日 2016-5-1
DOI 10.1093/mmy/myv106
PII myv106
PMID 26705837
MeSH DNA, Fungal / analysis DNA, Fungal / genetics Histoplasma / genetics* Histoplasmosis / microbiology* Humans Limit of Detection Molecular Typing / methods* Real-Time Polymerase Chain Reaction / methods*
IF 2.822
引用数 11
WOS 分野 VETERINARY SCIENCES MYCOLOGY INFECTIOUS DISEASES
リソース情報
病原微生物 NA