Development of a Multiple Detection Technique for Fungi by DNA Microarray with the Simultaneous Use of Internal Transcribed Spacer Region of Ribosomal RNA Gene and β-Tubulin Gene Probes

We offer the first description of the development of a multiple detection technique for fungi by DNA microarray with the simultaneous use of internal transcribed spacer region (ITS) of ribosomal RNA gene and β-tubulin gene probes. The assay uses 12 oligonucleotide probes and multiplex amplification...

Full description

Saved in:
Bibliographic Details
Published in:Biocontrol Science 2014, Vol.19(3), pp.139-145
Main Authors: ISSHIKI, ATSUNORI, TAKEHARU, HITOSHI, AOKI, SHUNSUKE, KOKAJI, MAMI, TANABE, SUGURU, KASETANI, TAISUKE, YOSHIDA, MITSUHIRO
Format: Article
Language:English
Subjects:
DNA microarray
Fungi
ITS region
Multiplex detection
β-tubulin gene
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We offer the first description of the development of a multiple detection technique for fungi by DNA microarray with the simultaneous use of internal transcribed spacer region (ITS) of ribosomal RNA gene and β-tubulin gene probes. The assay uses 12 oligonucleotide probes and multiplex amplification to detect fungal species belonging to various sections of Aspergillus, the Eurotium genus, and the Penicillium genus. The specificity of each probe was tested using 231 reference fungal strains, including 79 target and 152 non-target strains in 102 species of 24 genera. We determined the optimum concentration of the primer pairs for multiplex PCR to be 0.5 μM for the β-tubulin gene and 0.125 μM for the ITS region. In the field trial using 76 specimens containing 323 fungi (up to five fungal strains were included in one specimen), the concordance rate between the DNA microarray and the DNA sequencing results was 97.4％ at the species or genus levels.
ISSN:1342-4815
1884-0205
DOI:10.4265/bio.19.139