PMID- 26859628
OWN - NLM
STAT- MEDLINE
DCOM- 20170103
LR  - 20191210
IS  - 1556-7125 (Electronic)
IS  - 1535-3141 (Linking)
VI  - 13
IP  - 4
DP  - 2016 Apr
TI  - Real-Time TaqMan PCR Assay for the Detection of Heat-Labile and Heat-Stable 
      Enterotoxin Genes in a Geographically Diverse Collection of Enterotoxigenic 
      Escherichia coli Strains and Stool Specimens.
PG  - 212-20
LID - 10.1089/fpd.2015.2064 [doi]
AB  - Enterotoxigenic Escherichia coli (ETEC) are an important cause of diarrhea in 
      children under the age of 5 years in developing countries and are the leading 
      bacterial agent of traveler's diarrhea in persons traveling to these countries. 
      ETEC strains secrete heat-labile (LT) and/or heat-stable (ST) enterotoxins that 
      induce diarrhea by causing water and electrolyte imbalance. We describe the 
      validation of a real-time TaqMan PCR (RT-PCR) assay to detect LT, ST1a, and ST1b 
      enterotoxin genes in E. coli strains and in stool specimens. We validated LT/ST1b 
      duplex and ST1a single-plex RT-PCR assay using a conventional PCR assay as a gold 
      standard with 188 ETEC strains and 42 non-ETEC strains. We validated LT/ST1b 
      duplex and ST1a single-plex RT-PCR assay in stool specimens (n = 106) using 
      traditional culture as the gold standard. RT- PCR assay sensitivities for LT, 
      ST1a, and ST1b detection in strains were 100%, 100%, and 98%; specificities were 
      95%, 98%, and 99%, and Pearson correlation coefficient r was 0.9954 between 
      RT-PCR assay and the gold standard. In stool specimens, RT-PCR assay 
      sensitivities for LT, ST1a, and ST1b detection were 97%, 100%, and 97%; and 
      specificities were 99%, 94%, and 97%. Pearson correlation coefficient r was 
      0.9975 between RT-PCR results in stool specimens and the gold standard. Limits of 
      detection of LT, ST1a, and ST1b by RT-PCR assay were 0.1 to1.0 pg/muL and by 
      conventional PCR assay were 100 to1000 pg/muL. The accuracy, rapidity and 
      sensitivity of this RT-PCR assay is promising for ETEC detection in public 
      health/clinical laboratories and for laboratories in need of an independent 
      method to confirm results of other culture independent diagnostic tests.
FAU - Pattabiraman, Vaishnavi
AU  - Pattabiraman V
AD  - National Center for Zoonotic and Emerging Infectious Diseases, Centers for 
      Disease Control and Prevention , Atlanta, Georgia .
FAU - Parsons, Michele B
AU  - Parsons MB
AD  - National Center for Zoonotic and Emerging Infectious Diseases, Centers for 
      Disease Control and Prevention , Atlanta, Georgia .
FAU - Bopp, Cheryl A
AU  - Bopp CA
AD  - National Center for Zoonotic and Emerging Infectious Diseases, Centers for 
      Disease Control and Prevention , Atlanta, Georgia .
LA  - eng
PT  - Journal Article
PT  - Validation Study
DEP - 20160209
PL  - United States
TA  - Foodborne Pathog Dis
JT  - Foodborne pathogens and disease
JID - 101120121
RN  - 0 (DNA, Bacterial)
RN  - 0 (Enterotoxins)
RN  - 0 (Escherichia coli Proteins)
SB  - IM
MH  - Centers for Disease Control and Prevention, U.S.
MH  - DNA, Bacterial/isolation & purification/metabolism
MH  - Disease Outbreaks
MH  - Dysentery/*microbiology
MH  - Enterotoxigenic Escherichia coli/classification/*genetics/isolation & 
      purification/metabolism
MH  - Enterotoxins/chemistry/*genetics/metabolism
MH  - Epidemiological Monitoring
MH  - Escherichia coli Infections/diagnosis/epidemiology/*microbiology
MH  - Escherichia coli Proteins/chemistry/*genetics/metabolism
MH  - Feces/microbiology
MH  - Hot Temperature
MH  - Humans
MH  - Limit of Detection
MH  - Molecular Typing
MH  - Multiplex Polymerase Chain Reaction
MH  - Protein Stability
MH  - Real-Time Polymerase Chain Reaction
MH  - Reproducibility of Results
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sensitivity and Specificity
MH  - Time Factors
MH  - United States/epidemiology
EDAT- 2016/02/10 06:00
MHDA- 2017/01/04 06:00
CRDT- 2016/02/10 06:00
PHST- 2016/02/10 06:00 [entrez]
PHST- 2016/02/10 06:00 [pubmed]
PHST- 2017/01/04 06:00 [medline]
AID - 10.1089/fpd.2015.2064 [doi]
PST - ppublish
SO  - Foodborne Pathog Dis. 2016 Apr;13(4):212-20. doi: 10.1089/fpd.2015.2064. Epub 
      2016 Feb 9.