| RRC ID |
34454
|
| 著者 |
Isshiki A, Takeharu H, Aoki S, Kokaji M, Tanabe S, Kasetani T, Yoshida M.
|
| タイトル |
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.
|
| ジャーナル |
Biocontrol Sci
|
| Abstract |
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.
|
| 巻・号 |
19(3)
|
| ページ |
139-45
|
| 公開日 |
2014-1-1
|
| DOI |
10.4265/bio.19.139
|
| PII |
DN/JST.JSTAGE/bio/19.139
|
| PMID |
25252646
|
| MeSH |
Aspergillus / genetics
Aspergillus / isolation & purification*
DNA, Fungal / genetics
DNA, Ribosomal Spacer / genetics*
Eurotium / genetics
Eurotium / isolation & purification*
Microbiological Techniques / methods*
Molecular Diagnostic Techniques / methods
Oligonucleotide Array Sequence Analysis / methods*
Oligonucleotide Probes / genetics
Penicillium / genetics
Penicillium / isolation & purification*
Sensitivity and Specificity
Tubulin / genetics*
|
| IF |
0.833
|
| 引用数 |
2
|
|
WOS 分野
|
BIOLOGY
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| リソース情報 |
| 一般微生物 |
JCM 1575
JCM 10252
JCM 10253
JCM 22555
JCM 22971 |
