Publicação
Evaluation of Performances of Three DNA Enzyme Immunoassays for Detection of Helicobacter pylori PCR Products from Biopsy Specimens
| dc.contributor.author | Monteiro, Lurdes | |
| dc.contributor.author | Cabrita, José | |
| dc.contributor.author | Mégraud, Francis | |
| dc.date.accessioned | 2017-07-20T15:21:08Z | |
| dc.date.available | 2017-07-20T15:21:08Z | |
| dc.date.issued | 1997 | |
| dc.description.abstract | PCR is recognized as a promising method for the detection ofHeticobacterpytori in gastric biopsy specimens. However, detection of PCR products by gel electrophoresis is difficult to implement in routine clinical laboratories. The aim of this study was to compare three new DNA enzyme immunoassays with the standard method in their ability to detect PCR products. lhe three assays were based on Lhe amplification of a fragment of the ureC gene of H. pytori and a colorimetric hybridization assay. lhe first assay (GEN-ElI-K DNA enzyme immunoassay; Sorin, Sailugia, ltaly) was based on the hybridization of amplified DNA with a probe bound in microtiter wells and detected with labelled anti-DNA antibody. lhe second assay (Pylori-prob; Biocode, Sclessin, Belgium) comprised a solid-phase sandwich hybridization system with a specific biotinylated probe being used for detection. Finally, the third assay (PCR enzyme-linked immunosorbent assay; Boehringer, Mannheim, Germany) was based on the hybridization of amplified DNA Iabelled with digoxigenin as a probe (used as a coating in microtiter wells) and detected with antidigoxigenin-peroxidase as conjugate. The sensitivity of the colorimetric assay was evaluated by using amplification products from PCR assays performed on several 10-fotd dilutions of DNA from H. pylori CIP 101260, and the specificitv was assessed with dilferent urease-positive bacteria. Biopsy specimens from 199 patients were tested; 106 were classified as H. pylori positive, and 93 were classified as II. pylori negative by culture and/or histological examination as Lhe ‘goId standard.” lhe receiving operating characteristic curve was used to determine the best cutotf point for each assay. The detection of PCR products by colorimetric hybridization increases the sensitivit up to 100-told compared to that with gel electrophoresis. The results are rapid (4 h) and easy to interpret and can be automated. | pt_PT |
| dc.description.version | info:eu-repo/semantics/publishedVersion | pt_PT |
| dc.identifier.citation | Journal of Clinical Microbiology. 1997;35(11):2931-2936 | pt_PT |
| dc.identifier.issn | 0095-1137 | |
| dc.identifier.uri | http://hdl.handle.net/10451/28445 | |
| dc.language.iso | eng | pt_PT |
| dc.peerreviewed | yes | pt_PT |
| dc.relation.publisherversion | http://jcm.asm.org/content/35/11/2931.full.pdf+html | pt_PT |
| dc.title | Evaluation of Performances of Three DNA Enzyme Immunoassays for Detection of Helicobacter pylori PCR Products from Biopsy Specimens | pt_PT |
| dc.type | journal article | |
| dspace.entity.type | Publication | |
| oaire.citation.endPage | 2936 | pt_PT |
| oaire.citation.issue | 11 | pt_PT |
| oaire.citation.startPage | 2931 | pt_PT |
| oaire.citation.title | Journal of Clinical Microbiology | pt_PT |
| oaire.citation.volume | 35 | pt_PT |
| rcaap.rights | restrictedAccess | pt_PT |
| rcaap.type | article | pt_PT |