RRC ID 46816
著者 Sakai K, Trabasso P, Moretti ML, Mikami Y, Kamei K, Gonoi T.
タイトル Identification of fungal pathogens by visible microarray system in combination with isothermal gene amplification.
ジャーナル Mycopathologia
Abstract The increasing incidence of infectious diseases caused by fungi in immunocompromised patients has encouraged researchers to develop rapid and accurate diagnosis methods. Identification of the causative fungal species is critical in deciding the appropriate treatment, but it is not easy to get satisfactory results due to the difficulty of fungal cultivation and morphological identification from clinical samples. In this study, we established a microarray system that can identify 42 species from 24 genera of clinically important fungal pathogens by using a chemical color reaction in the detection process. The array uses the internal transcribed spacer region of the rRNA gene for identification of fungal DNA at the species level. The specificity of this array was tested against a total of 355 target and nontarget fungal species. The fungal detection was succeeded directly from 10(3) CFU/ml for whole blood samples, and 50 fg DNA per 1 ml of serum samples indicating that the array system we established is sensitive to identify infecting fungi from clinical sample. Furthermore, we conducted isothermal amplification in place of PCR amplification and labeling. The successful identification with PCR-amplified as well as isothermally amplified target genes demonstrated that our microarray system is an efficient and robust method for identifying a variety of fungal species in a sample.
巻・号 178(1-2)
ページ 11-26
公開日 2014-8-1
DOI 10.1007/s11046-014-9756-2
PMID 24952715
PMC PMC4098066
MeSH DNA, Fungal / genetics DNA, Ribosomal Spacer / genetics Fungi / classification* Fungi / genetics Fungi / isolation & purification* Humans Microarray Analysis / methods* Molecular Diagnostic Techniques / methods* Mycoses / diagnosis* Mycoses / microbiology Nucleic Acid Amplification Techniques / methods* Sensitivity and Specificity
IF 2.452
引用数 21
WOS 分野 MYCOLOGY
リソース情報
病原微生物 IFM 40718