| RRC ID |
21366
|
| 著者 |
Kurakawa T, Kubota H, Tsuji H, Matsuda K, Asahara T, Takahashi T, Ramamurthy T, Hamabata T, Takahashi E, Miyoshi S, Okamoto K, Mukhopadhyay AK, Takeda Y, Nomoto K.
|
| タイトル |
Development of a sensitive rRNA-targeted reverse transcription-quantitative polymerase chain reaction for detection of Vibrio cholerae/mimicus, V. parahaemolyticus/alginolyticus and Campylobacter jejuni/coli.
|
| ジャーナル |
Microbiol Immunol
|
| Abstract |
A sensitive rRNA-targeted reverse transcription-quantitative polymerase chain reaction (RT-qPCR) method was developed for detection of Vibrio cholerae/mimicus, V. parahaemolyticus/alginolyticus and Campylobacter jejuni/coli by using specific primers. Counts of the enteric pathogens spiked in human stools were quantified at the lower detection limit of 10(3) cells/g stool by RT-qPCR, in marked contrast with conventional quantitative polymerase chain reaction (qPCR) at the detection limit of 10(5) to 10(6) cells/g stool. The bacterial counts determined by RT-qPCR were almost equivalent to those determined by the culture method and fluorescence in situ hybridization (FISH) during the course of in vitro culture. Bacterial rRNA in the stools was stable for at least 4 weeks when the stools were kept as the suspensions in RNA-stabilizing agent, RNAlater®, even at 37(o) C. These data suggested that the rapid and high sensitive rRNA-targeted RT-qPCR was applicable for the accurate quantification of viable enteric pathogens, such as V. cholerae/mimicus, V. parahaemolyticus/alginolyticus and C. jejuni/coli.
|
| 巻・号 |
56(1)
|
| ページ |
10-20
|
| 公開日 |
2012-1-1
|
| DOI |
10.1111/j.1348-0421.2011.00405.x
|
| PMID |
22146006
|
| MeSH |
Bacterial Load / methods
Campylobacter coli / isolation & purification*
Campylobacter jejuni / isolation & purification
DNA Primers / genetics
Feces / microbiology
Genes, rRNA
Humans
In Situ Hybridization, Fluorescence
RNA, Bacterial / genetics
RNA, Ribosomal, 16S / genetics
Reverse Transcriptase Polymerase Chain Reaction / methods*
Sensitivity and Specificity
Specimen Handling
Vibrio cholerae / isolation & purification
Vibrio mimicus / isolation & purification
Vibrio parahaemolyticus / isolation & purification*
|
| IF |
1.566
|
| 引用数 |
17
|
|
WOS 分野
|
MICROBIOLOGY
IMMUNOLOGY
|
| リソース情報 |
| 一般微生物 |
JCM 1027
JCM 1043
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JCM 1232
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JCM 1665
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JCM 1668
JCM 1669
JCM 1672
JCM 1673
JCM 2013
JCM 2152
JCM 2527
JCM 2530
JCM 3752
JCM 3943
JCM 3948
JCM 5802
JCM 5824
JCM 6301
JCM 7375 |
