Evaluation of DNA-DNA hybridization for the direct detection of enterotoxigenic Escherichia coli in stool blots.

The simplicity of enterotoxigenic Escherichia coli (ETEC) stool blot hybridization, where the total bacterial growth of a fecal inoculum is examined directly for the presence of enterotoxin genes, has been marred by reports of unsatisfactory sensitivity and/or specificity. To assess the accuracy of stool blot hybridization and to study the effect of varying proportions of ETEC among fecal E. coli (ETEC/E. coli) on test performance, a detailed 'blind' study of 166 stool specimens from children with diarrhea was performed. Oligonucleotide probes were found to be superior to polynucleotide probes, having a sensitivity of 80%, a specificity of 99%, a positive predictive value of 89% and a negative predictive value of 97%. The sensitivity was found to be 100% when ETEC/E. coli > 2/12, as compared with 20% when ETEC/E. coli < or = 2/12 (p = 0.001), showing that the proportion of ETEC among fecal E. coli is of paramount importance for test sensitivity.

[1]  M. Bhan,et al.  Use of nonradioactive DNA hybridization for identification of enterotoxigenic Escherichia coli harboring genes for colonization factor antigen I, coli surface antigen 4, or putative colonization factor O166 , 1992, Journal of clinical microbiology.

[2]  M. Bhan,et al.  Simplified and accurate nonradioactive polynucleotide gene probe assay for identification of enterotoxigenic Escherichia coli , 1990, Journal of clinical microbiology.

[3]  G. Schoolnik,et al.  Multi‐gene amplification: simultaneous detection of three virulence genes in diarrhoeal stool , 1989, Molecular microbiology.

[4]  R. Black,et al.  Incidence and etiology of infantile diarrhea and major routes of transmission in Huascar, Peru. , 1989, American journal of epidemiology.

[5]  J. Brown,et al.  Examination of colonies and stool blots for detection of enteropathogens by DNA hybridization with eight DNA probes , 1989, Journal of clinical microbiology.

[6]  D. M. Olive,et al.  Detection of enterotoxigenic Escherichia coli after polymerase chain reaction amplification with a thermostable DNA polymerase , 1989, Journal of clinical microbiology.

[7]  H. Sommerfelt,et al.  Oligonucleotide gene probes may detect enterotoxigenic Escherichia coli directly in fecal smears , 1989, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[8]  K. Kalland,et al.  Cloned polynucleotide and synthetic oligonucleotide probes used in colony hybridization are equally efficient in the identification of enterotoxigenic Escherichia coli , 1988, Journal of clinical microbiology.

[9]  K. Kalland,et al.  Comparative study of colony hybridization with synthetic oligonucleotide probes and enzyme-linked immunosorbent assay for identification of enterotoxigenic Escherichia coli , 1988, Journal of clinical microbiology.

[10]  M. Yoshikawa,et al.  Detection of enterotoxigenic Escherichia coli by colony hybridization with biotinylated enterotoxin probes , 1987, Journal of clinical microbiology.

[11]  H. Dupont,et al.  Utility of Oligo deoxyribonucleotide Probes for Detecting Enterotoxigenic Escherichia coli , 1987 .

[12]  J. Holmgren,et al.  Monoclonal antibodies against Escherichia coli heat-stable toxin (STa) and their use in a diagnostic ST ganglioside GM1-enzyme-linked immunosorbent assay , 1986, Journal of clinical microbiology.

[13]  M. Lung,et al.  Evaluation and optimization of the DNA filter assay for direct detection of enterotoxigenic Escherichia coli in the presence of stool coliforms , 1986, Journal of clinical microbiology.

[14]  P. Echeverria,et al.  A comparative study of enterotoxin gene probes and tests for toxin production to detect enterotoxigenic Escherichia coli. , 1986, The Journal of infectious diseases.

[15]  J. Holmgren,et al.  Monoclonal Antibodies and Immunodetection Methods for Vibrio cholerae and Escherichia coli Enterotoxins , 1986 .

[16]  M. Levine,et al.  Sensitivity and specificity of DNA probes with the stool blot technique for detection of Escherichia coli enterotoxins. , 1985, The Journal of infectious diseases.

[17]  W. L. Payne,et al.  Synthetic oligodeoxyribonucleotide probes for detecting heat-stable enterotoxin-producing Escherichia coli by DNA colony hybridization , 1985, Applied and environmental microbiology.

[18]  David N. Taylor,et al.  Identification by DNA hybridization of enterotoxigenic Escherichia coli in a longitudinal study of villages in Thailand. , 1985, The Journal of infectious diseases.

[19]  H. Brandwein,et al.  Simple and reliable enzyme-linked immunosorbent assay with monoclonal antibodies for detection of Escherichia coli heat-stable enterotoxins , 1984, Journal of clinical microbiology.

[20]  R. Holmes,et al.  Characterization of monoclonal antibodies to heat-labile enterotoxin encoded by a plasmid from a clinical isolate of Escherichia coli , 1984, Infection and immunity.

[21]  P. Echeverria,et al.  Isolation and nucleotide sequence determination of a gene encoding a heat-stable enterotoxin of Escherichia coli , 1983, Infection and immunity.

[22]  P. Echeverria,et al.  Identification of enterotoxigenic Escherichia coli by colony hybridization using three enterotoxin gene probes. , 1982, The Journal of infectious diseases.

[23]  S. Falkow,et al.  Detection of enterotoxigenic Escherichia coli by DNA colony hybridization. , 1980, The Journal of infectious diseases.

[24]  J. Shaffer,et al.  Hybridization of synthetic oligodeoxyribonucleotides to ΦX 174 DNA: the effect of single base pair mismatch , 1979 .

[25]  R. Sack,et al.  Use of colony pools for diagnosis of enterotoxigenic Escherichia coli diarrhea , 1979, Journal of clinical microbiology.

[26]  R. Giannella.  Suckling mouse model for detection of heat-stable Escherichia coli enterotoxin: characteristics of the model , 1976, Infection and immunity.

[27]  D. Sack,et al.  Test for enterotoxigenic Escherichia coli using Y-1 adrenal cells in miniculture , 1975, Infection and immunity.

[28]  A. Gilman,et al.  Cyclic Adenosine Monophosphate and Alteration of Chinese Hamster Ovary Cell Morphology: a Rapid, Sensitive In Vitro Assay for the Enterotoxins of Vibrio cholerae and Escherichia coli , 1974, Infection and immunity.