title: Identification of fungal pathogens by visible microarray system in combination with isothermal gene amplification.
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.
authors: Division of BioResources, Medical Mycology Research Center, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba, Japan.
authors: Kanae
authors: K
authors: Sakai
authors: Plinio
authors: P
authors: Trabasso
authors: Maria Luiza
authors: ML
authors: Moretti
authors: Yuzuru
authors: Y
authors: Mikami
authors: Katsuhiko
authors: K
authors: Kamei
authors: Tohru
authors: T
authors: Gonoi
mesh_terms: D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005658:Fungi / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D046228:Microarray Analysis / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009181:Mycoses / Q000175:diagnosis* / Q000382:microbiology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer