title: Detection of PCR products of the ipaH gene from Shigella and enteroinvasive Escherichia coli by enzyme linked immunosorbent assay.
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 (SiO(2)). 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.
authors.affiliation: Department of Enteric Diseases, Armed Forces Research Institute of Medical Science, Bangkok, Thailand.
authors.forename: O
authors.initials: O
authors.lastname: Sethabutr
authors.forename: M
authors.initials: M
authors.lastname: Venkatesan
authors.forename: S
authors.initials: S
authors.lastname: Yam
authors.forename: L W
authors.initials: LW
authors.lastname: Pang
authors.forename: B L
authors.initials: BL
authors.lastname: Smoak
authors.forename: W K
authors.initials: WK
authors.lastname: Sang
authors.forename: P
authors.initials: P
authors.lastname: Echeverria
authors.forename: D N
authors.initials: DN
authors.lastname: Taylor
authors.forename: D W
authors.initials: DW
authors.lastname: Isenbarger
mesh_terms: D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000235:genetics*; D002498:Centrifugation; D002648:Child; D002675:Child, Preschool; D003967:Diarrhea / Q000382:microbiology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D005798:Genes, Bacterial; D006801:Humans; D007223:Infant; D016133:Polymerase Chain Reaction / Q000379:methods; D011791:Quartz; D012760:Shigella / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C063500:ipaH protein, Shigella flexneri; D011791:Quartz