Detection of PCR products of the ipaH gene from Shigella and enteroinvasive Escherichia coli by enzyme linked immunosorbent assay

Orntipa Sethabutr, Malabi Venkatesan, Sylvia Yam, Lorrin W. Pang, Bonnie L. Smoak, Willie K. Sang, Peter Echeverria, David N. Taylor, Daniel W. Isenbarger

Research output: Contribution to journalArticle

Abstract

PCR techniques applied to diarrheal stools reliably diagnose Shigella and enteroinvasive Escherichia coli (EIEC) infections. Identification of PCR products using agarose gel electrophoresis (AGE) and hybridization with DNA probes has several shortcomings. Automated methods of identifying PCR products that process larger numbers of specimens can facilitate epidemiologic studies and standardize results. In this study, we used ELISA following PCR to detect ipaH gene sequences of Shigella and EIEC from 89 diarrheal stools. Results of ELISA were compared with AGE with and without DNA probe, and with culture. Two specimen preparation methods were compared as well: boiling/centrifugation, and purification with silicon dioxide (SiO2). Both PCR product-detection methods identified significantly more infections than did culture. PCR-ELISA detected significantly more infections than PCR-AGE when processed using SiO2 (P = 0.014). PCR-ELISA allows screening of larger numbers of specimens, automates test results, and avoids use of mutagenic reagents. PCR-ELISA is faster than PCR-AGE when testing large numbers of specimens, although not when testing small numbers of specimens. Copyright (C) 2000 Elsevier Science Inc.

Original languageEnglish (US)
Pages (from-to)11-16
Number of pages6
JournalDiagnostic Microbiology and Infectious Disease
Volume37
Issue number1
DOIs
StatePublished - May 2000
Externally publishedYes

Fingerprint

Shigella
Enzyme-Linked Immunosorbent Assay
Escherichia coli
Polymerase Chain Reaction
Agar Gel Electrophoresis
Genes
DNA Probes
Escherichia coli Infections
Infection
Centrifugation
Silicon Dioxide
Epidemiologic Studies

Keywords

  • Diagnosis
  • EIEC
  • ELISA
  • ipaH
  • PCR
  • Shigella

ASJC Scopus subject areas

  • Infectious Diseases
  • Immunology and Allergy
  • Virology
  • Parasitology
  • Microbiology
  • Immunology
  • Applied Microbiology and Biotechnology

Cite this

Detection of PCR products of the ipaH gene from Shigella and enteroinvasive Escherichia coli by enzyme linked immunosorbent assay. / Sethabutr, Orntipa; Venkatesan, Malabi; Yam, Sylvia; Pang, Lorrin W.; Smoak, Bonnie L.; Sang, Willie K.; Echeverria, Peter; Taylor, David N.; Isenbarger, Daniel W.

In: Diagnostic Microbiology and Infectious Disease, Vol. 37, No. 1, 05.2000, p. 11-16.

Research output: Contribution to journalArticle

Sethabutr, O, Venkatesan, M, Yam, S, Pang, LW, Smoak, BL, Sang, WK, Echeverria, P, Taylor, DN & Isenbarger, DW 2000, 'Detection of PCR products of the ipaH gene from Shigella and enteroinvasive Escherichia coli by enzyme linked immunosorbent assay', Diagnostic Microbiology and Infectious Disease, vol. 37, no. 1, pp. 11-16. https://doi.org/10.1016/S0732-8893(00)00122-X
Sethabutr, Orntipa ; Venkatesan, Malabi ; Yam, Sylvia ; Pang, Lorrin W. ; Smoak, Bonnie L. ; Sang, Willie K. ; Echeverria, Peter ; Taylor, David N. ; Isenbarger, Daniel W. / Detection of PCR products of the ipaH gene from Shigella and enteroinvasive Escherichia coli by enzyme linked immunosorbent assay. In: Diagnostic Microbiology and Infectious Disease. 2000 ; Vol. 37, No. 1. pp. 11-16.
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