Comparison of one commercial and two in-house TaqMan multiplex real-time PCR assays for detection of enteropathogenic, enterotoxigenic and enteroaggregative Escherichia coli
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Comparison of one commercial and two in-house TaqMan multiplex real-time PCR assays for detection of enteropathogenic, enterotoxigenic and enteroaggregative Escherichia coli. / Hahn, Andreas; Lütgehetmann, Marc; Landt, Olfert; Schwarz, Norbert Georg; Frickmann, Hagen.
in: TROP MED INT HEALTH, Jahrgang 22, Nr. 11, 11.2017, S. 1371-1376.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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T1 - Comparison of one commercial and two in-house TaqMan multiplex real-time PCR assays for detection of enteropathogenic, enterotoxigenic and enteroaggregative Escherichia coli
AU - Hahn, Andreas
AU - Lütgehetmann, Marc
AU - Landt, Olfert
AU - Schwarz, Norbert Georg
AU - Frickmann, Hagen
N1 - © 2017 John Wiley & Sons Ltd.
PY - 2017/11
Y1 - 2017/11
N2 - OBJECTIVE: Enteropathogenic, enterotoxigenic and enteroaggregative Escherichia coli (EPEC, ETEC, EAEC) are among the most frequent causes of diarrhoea during travel or on military deployments. Cost-efficient and reliable real-time multiplex PCR (mPCR) assays are desirable for surveillance or point prevalence studies in remote and resource-limited tropical settings. We compared one commercial PCR kit and two in-house assays without using a gold standard to estimate sensitivity and specificity of each assay.METHODS: Residual materials from nucleic acid extractions of stool samples from two groups with presumably different prevalences and increased likelihood of being infected or colonised by diarrhoeagenic E. coli were included in the assessment. One group comprised samples from returnees from tropical deployments, the second group was of migrants and study participants from high-endemicity settings. Each sample was assessed with all of the PCR assays. Cycle threshold (Ct) values were descriptively compared.RESULTS: The calculated sensitivities for the commercial test vs. the in-house tests were for EPEC 0.84 vs. 0.89 and 0.96, for ETEC 0.83 vs. 0.76 and 0.61, and for EAEC 0.69 vs. 0.54 and 0.69. False positive results were rare - specificity was 0.94 and 0.97 for two EPEC tests and 1.0 for all other tests. Most positive samples had late Ct values corresponding to low quantities of pathogens. Discordant test results were associated with late Ct values.CONCLUSIONS: As commercial and in-house assays showed comparable results, in-house tests can be assumed to be safe while affording considerable savings, making them a valuable alternative for surveillance testing in resource-limited tropical areas.
AB - OBJECTIVE: Enteropathogenic, enterotoxigenic and enteroaggregative Escherichia coli (EPEC, ETEC, EAEC) are among the most frequent causes of diarrhoea during travel or on military deployments. Cost-efficient and reliable real-time multiplex PCR (mPCR) assays are desirable for surveillance or point prevalence studies in remote and resource-limited tropical settings. We compared one commercial PCR kit and two in-house assays without using a gold standard to estimate sensitivity and specificity of each assay.METHODS: Residual materials from nucleic acid extractions of stool samples from two groups with presumably different prevalences and increased likelihood of being infected or colonised by diarrhoeagenic E. coli were included in the assessment. One group comprised samples from returnees from tropical deployments, the second group was of migrants and study participants from high-endemicity settings. Each sample was assessed with all of the PCR assays. Cycle threshold (Ct) values were descriptively compared.RESULTS: The calculated sensitivities for the commercial test vs. the in-house tests were for EPEC 0.84 vs. 0.89 and 0.96, for ETEC 0.83 vs. 0.76 and 0.61, and for EAEC 0.69 vs. 0.54 and 0.69. False positive results were rare - specificity was 0.94 and 0.97 for two EPEC tests and 1.0 for all other tests. Most positive samples had late Ct values corresponding to low quantities of pathogens. Discordant test results were associated with late Ct values.CONCLUSIONS: As commercial and in-house assays showed comparable results, in-house tests can be assumed to be safe while affording considerable savings, making them a valuable alternative for surveillance testing in resource-limited tropical areas.
KW - Commerce
KW - Diarrhea
KW - Endemic Diseases
KW - Escherichia coli
KW - Escherichia coli Infections
KW - Feces
KW - Health Resources
KW - Humans
KW - Military Personnel
KW - Multiplex Polymerase Chain Reaction
KW - Real-Time Polymerase Chain Reaction
KW - Sensitivity and Specificity
KW - Transients and Migrants
KW - Travel
KW - Tropical Climate
KW - Comparative Study
KW - Evaluation Studies
KW - Journal Article
U2 - 10.1111/tmi.12976
DO - 10.1111/tmi.12976
M3 - SCORING: Journal article
C2 - 28906580
VL - 22
SP - 1371
EP - 1376
JO - TROP MED INT HEALTH
JF - TROP MED INT HEALTH
SN - 1360-2276
IS - 11
ER -