PMID- 24048535
OWN - NLM
STAT- MEDLINE
DCOM- 20140623
LR  - 20211021
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Print)
IS  - 0095-1137 (Linking)
VI  - 51
IP  - 12
DP  - 2013 Dec
TI  - Quantitative detection of Vibrio cholera toxin by real-time and dynamic 
      cytotoxicity monitoring.
PG  - 3968-74
LID - 10.1128/JCM.01959-13 [doi]
AB  - We report here the quantitative detection of Vibrio cholerae toxin (CT) in 
      isolates and stool specimens by dynamic monitoring of the full course of 
      CT-mediated cytotoxicity in a real-time cell analysis (RTCA) system. Four cell 
      lines, including Y-1 mouse adrenal tumor cells, Chinese hamster ovary (CHO) 
      cells, small intestine epithelial (FHs74Int) cells, and mouse adrenal gland 
      (PC12-Adh) cells, were evaluated for their suitability for CT-induced 
      cytotoxicity testing. Among them, the Y-1 line was demonstrated to be the most 
      sensitive for CT-mediated cytotoxicity, with limits of detection of 7.0 pg/ml for 
      purified CT and 0.11 ng/ml for spiked CT in pooled negative stool specimens. No 
      CT-mediated cytotoxicity was observed for nontoxigenic V. cholerae, non-V. 
      cholerae species, or non-V. cholerae enterotoxins. The CT-RTCA assay was further 
      validated with 100 stool specimens consecutively collected from patients with 
      diarrhea and 200 V. cholerae isolates recovered from patients and the 
      environment, in comparison to a reference using three detection methods. The 
      CT-RTCA assay had sensitivities and specificities of 97.5% and 100.0%, 
      respectively, for V. cholerae isolates and 90.0% and 97.2% for stool specimens. 
      For stool specimens spiked with CT concentrations ranging from 3.5 pg/ml to 1.8 
      ng/ml, the inoculation-to-detection time was 1.12 +/- 0.38 h, and the values were 
      inversely correlated with CT concentrations (rho = -1; P = 0.01). The results 
      indicate that the CT-RTCA assay with the Y-1 cell line provides a rapid and 
      sensitive tool for the quantitative detection of CT activities in clinical 
      specimens.
FAU - Jin, Dazhi
AU  - Jin D
AD  - Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, China.
FAU - Luo, Yun
AU  - Luo Y
FAU - Zheng, Min
AU  - Zheng M
FAU - Li, Haijing
AU  - Li H
FAU - Zhang, Jing
AU  - Zhang J
FAU - Stampfl, Melinda
AU  - Stampfl M
FAU - Xu, Xiao
AU  - Xu X
FAU - Ding, Gangqiang
AU  - Ding G
FAU - Zhang, Yanjun
AU  - Zhang Y
FAU - Tang, Yi-Wei
AU  - Tang YW
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130918
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 9012-63-9 (Cholera Toxin)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cell Survival/drug effects
MH  - Cholera/*diagnosis
MH  - Cholera Toxin/*analysis/*toxicity
MH  - Cricetinae
MH  - Cricetulus
MH  - Diarrhea/diagnosis
MH  - Humans
MH  - Mice
MH  - Sensitivity and Specificity
MH  - Vibrio cholerae/*metabolism
PMC - PMC3838090
EDAT- 2013/09/21 06:00
MHDA- 2014/06/24 06:00
PMCR- 2014/06/01
CRDT- 2013/09/20 06:00
PHST- 2013/09/20 06:00 [entrez]
PHST- 2013/09/21 06:00 [pubmed]
PHST- 2014/06/24 06:00 [medline]
PHST- 2014/06/01 00:00 [pmc-release]
AID - JCM.01959-13 [pii]
AID - 01959-13 [pii]
AID - 10.1128/JCM.01959-13 [doi]
PST - ppublish
SO  - J Clin Microbiol. 2013 Dec;51(12):3968-74. doi: 10.1128/JCM.01959-13. Epub 2013 
      Sep 18.