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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
Author(s) -
Hahn Andreas,
Luetgehetmann Marc,
Landt Olfert,
Schwarz Norbert Georg,
Frickmann Hagen
Publication year - 2017
Publication title -
tropical medicine and international health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.056
H-Index - 114
eISSN - 1365-3156
pISSN - 1360-2276
DOI - 10.1111/tmi.12976
Subject(s) - taqman , gold standard (test) , multiplex , microbiology and biotechnology , multiplex polymerase chain reaction , virology , biology , enterotoxigenic escherichia coli , polymerase chain reaction , medicine , veterinary medicine , escherichia coli , enterotoxin , bioinformatics , biochemistry , gene
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.