PMID- 22146006
OWN - NLM
STAT- MEDLINE
DCOM- 20120530
LR  - 20120214
IS  - 1348-0421 (Electronic)
IS  - 0385-5600 (Linking)
VI  - 56
IP  - 1
DP  - 2012 Jan
TI  - 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.
PG  - 10-20
LID - 10.1111/j.1348-0421.2011.00405.x [doi]
AB  - 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(R), 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.
CI  - (c) 2012 The Societies and Blackwell Publishing Asia Pty Ltd.
FAU - Kurakawa, Takashi
AU  - Kurakawa T
AD  - Yakult Central Institute for Microbiological Research, 1796 Yaho, Kunitachi, 
      Tokyo 186-8650, Japan. takashi-kurakawa@yakult.co.jp
FAU - Kubota, Hiroyuki
AU  - Kubota H
FAU - Tsuji, Hirokazu
AU  - Tsuji H
FAU - Matsuda, Kazunori
AU  - Matsuda K
FAU - Asahara, Takashi
AU  - Asahara T
FAU - Takahashi, Takuya
AU  - Takahashi T
FAU - Ramamurthy, Thandavarayan
AU  - Ramamurthy T
FAU - Hamabata, Takashi
AU  - Hamabata T
FAU - Takahashi, Eizo
AU  - Takahashi E
FAU - Miyoshi, Shin-ichi
AU  - Miyoshi S
FAU - Okamoto, Keinosuke
AU  - Okamoto K
FAU - Mukhopadhyay, Asish K
AU  - Mukhopadhyay AK
FAU - Takeda, Yoshifumi
AU  - Takeda Y
FAU - Nomoto, Koji
AU  - Nomoto K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Australia
TA  - Microbiol Immunol
JT  - Microbiology and immunology
JID - 7703966
RN  - 0 (DNA Primers)
RN  - 0 (RNA, Bacterial)
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Bacterial Load/methods
MH  - Campylobacter coli/*isolation & purification
MH  - Campylobacter jejuni/isolation & purification
MH  - DNA Primers/genetics
MH  - Feces/microbiology
MH  - Genes, rRNA
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - RNA, Bacterial/genetics
MH  - RNA, Ribosomal, 16S/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
MH  - Sensitivity and Specificity
MH  - Specimen Handling
MH  - Vibrio cholerae/isolation & purification
MH  - Vibrio mimicus/isolation & purification
MH  - Vibrio parahaemolyticus/*isolation & purification
EDAT- 2011/12/08 06:00
MHDA- 2012/05/31 06:00
CRDT- 2011/12/08 06:00
PHST- 2011/12/08 06:00 [entrez]
PHST- 2011/12/08 06:00 [pubmed]
PHST- 2012/05/31 06:00 [medline]
AID - 10.1111/j.1348-0421.2011.00405.x [doi]
PST - ppublish
SO  - Microbiol Immunol. 2012 Jan;56(1):10-20. doi: 10.1111/j.1348-0421.2011.00405.x.