Detection of Diarrheogenic Escherichia coli Using Nucleotide Probes

Echeverria, Peter; Seriwatana, Jitvimol; Sethabutr, Orntipa; Chatkaeomorakot, Arunsri

doi:10.1016/b978-0-12-463000-0.50010-4

Cited by 29 publications

(19 citation statements)

References 156 publications

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“…These Dig-labeled DNA probes were used for detecting genes for human heat-labile enterotoxin (elth); human heat-stable enterotoxin (esth); porcine heatstable enterotoxin (estp); shiga toxin 1 (stx1); shiga-like toxin type2e (stx2e); invasion-associated locus of the invasion plasmid in EIEC (ial); bundle-forming pilin A (bfpA); enteroadherent factor plasmid (eaf), effacing and attachment factor (eae); and aggregative adherent factors (aaf), using the standard operating procedures of the Department of Enteric Diseases of the Armed Forces Research Institute of Medical Science, Thailand [2,9]. Briefly, the bacterial colonies were spotted and cultured on an N+ nylon membrane (Amersham, Braunschweig, Germany) on dry Trypticase Soy agar plates.…”

Section: Methodsmentioning

confidence: 99%

Virulence Genes and Antimicrobial Susceptibilities of Hemolytic and Nonhemolytic Escherichia coli Isolated from Post-Weaning Piglets in Central Thailand

Prapasarakul

Tummaruk

Niyomtum

et al. 2010

The Journal of Veterinary Medical Science

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ABSTRACT. The purpose of this study was to compare the existence of virulence genes in hemolytic Escherichia coli (HEC) and nonhemolytic E. coli (NHEC) isolated from weaner pigs in Thailand, and to determine their susceptibility to 10 antimicrobial agents. A total of 304 E. coli isolates were obtained from 90 piglets with diarrhea and 110 healthy piglets. Of these, 74 HEC isolates were obtained from 70 pigs with diarrhea, and 4 were obtained from 4 healthy pigs, while 190 and 40 NHEC were recovered from 110 healthy and 20 pigs with diarrhea, respectively. A ten digoxigenin (DIG)-labeled probe system was utilized for detecting genes encoding virulence-associated toxins and proteins in these isolates, and the minimal inhibitory concentration values against 10 antimicrobials were determined by means of the agar dilution technique. In total, 70.3% of the HEC isolates contained an exotoxin gene, lth, estp or stx2e, whereas 2.6% of the NHEC isolates hybridized with a gene probe for estp or stx2e. Over 90% of the isolates were resistant to most agents other than colistin and halquinol. The MIC 90 values of the HEC isolates for halquinol and colistin were 4 and 8 times greater than those of the NHEC isolates, respectively. The results represent the first characterization of resistant pathogenic E. coli distributed in the Thai pig industry. Amongst the HEC isolates, there appeared to be an association between the presence of some exotoxin genes, including lth, estp and stx2e, and reduced antimicrobial susceptibility.

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Section: Methodsmentioning

confidence: 99%

Virulence Genes and Antimicrobial Susceptibilities of Hemolytic and Nonhemolytic Escherichia coli Isolated from Post-Weaning Piglets in Central Thailand

Prapasarakul

Tummaruk

Niyomtum

et al. 2010

The Journal of Veterinary Medical Science

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“…6,7,8 Five lactose-fermenting and five non-lactose-fermenting E. coli colonies per specimen were tested using DNA probes for detection of ETEC toxins, enteroinvasive E. coli, enterohemorrhagic E. coli, locally adherent enteropathogenic E. coli, and attaching and effacing E. coli (eae+ E. coli). 9,10 Stool specimens were examined for the presence of rotavirus by a commercially available enzyme-linked immunosorbent assay (Rotazyme; Abbott Laboratories, North Chicago, IL). A reverse transcription−polymerase chain reaction assay was performed as previously described to detect the presence of Norwalk virus.…”

Section: Methodsmentioning

confidence: 99%

An observational clinic-based study of diarrheal illness in deployed United States military personnel in Thailand: presentation and outcome of Campylobacter infection.

Sanders¹,

Isenbarger²,

Walz³

et al. 2002

The American Journal of Tropical Medicine and Hygiene

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Abstract. Campylobacter is a leading cause of traveler's diarrhea in Thailand. Since resistance to quinolones is high among Campylobacter isolates, empiric therapy with quinolones for traveler's diarrhea may be ineffective in this region. We conducted an observational study among 169 U.S. military personnel with acute diarrhea and compared their microbiologic findings to those of 77 asymptomatic personnel deployed to Thailand in May 1998. Of 146 pathogenic bacterial isolates, the most common were nontyphoidal Salmonella (n ‫ס‬ 31), enterotoxigenic Escherichia coli (n ‫ס‬ 24), and C. jejuni/coli (n ‫ס‬ 23). Campylobacter was strongly associated with disease (odds ratio ‫ס‬ 5.9; 95% confidence interval ‫ס‬ 1.3−37.3), with a more severe clinical presentation, and with a reduced functional ability at presentation (P ‫ס‬ 0.02). In vitro resistance to ciprofloxacin was observed in 96% of the Campylobacter isolates. Sub-optimal treatment response to ciprofloxacin was observed in 17% of the cases of Campylobacter infection versus 6% due to other causes. These results highlight the importance of Campylobacter as a cause of severe traveler's diarrhea in Thailand and illustrates the ongoing problem with antibiotic-resistant strains and associated treatment problems.

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“…7 Shiga-like toxin I and II (SLTI and SLT II) probes were used to identify Shiga-like toxin-producing E. coli (STEC). 7 Effacing attachment (EAE), Enteroadherent factor (EAF), and bundle-forming pilus A (bfpA) probes were used to identify Enteropathogenic E. coli (EPEC). 8,9 Three reference E. coli strains derived from plasmid harboring enterotoxin genes, pEWD299 (LT), pDAS100 (STIa), and pDAS101 (STIb), were included as positive controls for ETEC.…”

Section: Introductionmentioning

confidence: 99%

“…A heat-labile toxin (LT) and two heat-stable toxin (STIa and STIb) probes were used to identify Enterotoxigenic E. coli (ETEC). 6,7 Invasion-associated locus (ial) probe was used to identify Enteroinvasive E. coli (EIEC). 7 Shiga-like toxin I and II (SLTI and SLT II) probes were used to identify Shiga-like toxin-producing E. coli (STEC).…”

Section: Introductionmentioning

confidence: 99%

“…6,7 Invasion-associated locus (ial) probe was used to identify Enteroinvasive E. coli (EIEC). 7 Shiga-like toxin I and II (SLTI and SLT II) probes were used to identify Shiga-like toxin-producing E. coli (STEC). 7 Effacing attachment (EAE), Enteroadherent factor (EAF), and bundle-forming pilus A (bfpA) probes were used to identify Enteropathogenic E. coli (EPEC).…”

Section: Introductionmentioning

confidence: 99%

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Case-Control Study of Diarrheal Disease Etiology in a Remote Rural Area in Western Thailand

Bodhidatta¹,

McDaniel²,

Sornsakrin³

et al. 2010

The American Society of Tropical Medicine and Hygiene

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Detection of Diarrheogenic Escherichia coli Using Nucleotide Probes

Cited by 29 publications

References 156 publications

Virulence Genes and Antimicrobial Susceptibilities of Hemolytic and Nonhemolytic Escherichia coli Isolated from Post-Weaning Piglets in Central Thailand

Virulence Genes and Antimicrobial Susceptibilities of Hemolytic and Nonhemolytic Escherichia coli Isolated from Post-Weaning Piglets in Central Thailand

An observational clinic-based study of diarrheal illness in deployed United States military personnel in Thailand: presentation and outcome of Campylobacter infection.

Case-Control Study of Diarrheal Disease Etiology in a Remote Rural Area in Western Thailand

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