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Detection of Helicobacter pylori by enzyme-linked immunosorbent assay of thermophilic helicase-dependent isothermal DNA amplification

Gill, P. and Amini, M. and Ghaemi, A. and Shokouhizadeh, L. and Abdul-Tehrani, H. and Karami, A. and Gilak, A. (2007) Detection of Helicobacter pylori by enzyme-linked immunosorbent assay of thermophilic helicase-dependent isothermal DNA amplification. Diagnostic Microbiology and Infectious Disease, 59 (3). pp. 243-249. ISSN 07328893 (ISSN)

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Abstract

An enzyme-linked immunosorbent assay (ELISA) of thermophilic helicase-dependent isothermal DNA amplification (tHDA) was developed for detection of Helicobacter pylori. The primers targeting ureC were used for the amplification of bacterial DNA by the isothermal digoxigenin (DIG)-labeling tHDA process, resulting in the accumulation of DIG-labeled DNA amplicons. The amplicons were denatured using heat and then hybridized with a specific biotinylated DNA probe, which was noncovalently immobilized on streptavidin-coated microtiter plate. The hybrids were colorimetrically detected by the addition of an anti-DIG antibody HRP conjugate and 2,2-azino-di-(3-ethylbenzthiazolinsulfonate) substrate solution. Results obtained from the gastric biopsy samples showed 90% and 95.7% of sensitivity and specificity, respectively, in comparison with culture results, and 96.6% and 96.8% of sensitivity and specificity, respectively, in comparison with those of the histologic studies. This assay significantly reduces the time needed for the identification of H. pylori and has the potential to facilitate early detection of this gastrointestinal pathogen. © 2007 Elsevier Inc. All rights reserved.

Item Type: Article
Additional Information: Unmapped bibliographic data: LA - English [Field not mapped to EPrints] J2 - Diagn. Microbiol. Infect. Dis. [Field not mapped to EPrints] C2 - 17662567 [Field not mapped to EPrints] AD - Research Center for Molecular Biology, Baqiyatallah Medical Sciences University, Tehran, 16739-78964, Iran [Field not mapped to EPrints] AD - Baqiyatallah Hospital, Tehran, 14359-15371, Iran [Field not mapped to EPrints] AD - Faculty of Medicine, Golestan University of Medical Sciences and Health Care, P.O. Box 619, Gorgan, Iran [Field not mapped to EPrints] AD - Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, 14115-331, Iran [Field not mapped to EPrints] AD - Department of Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, 14115-331, Iran [Field not mapped to EPrints] AD - ISOCHEM, 53340 Meckenheim, Germany [Field not mapped to EPrints] DB - Scopus [Field not mapped to EPrints]
Uncontrolled Keywords: Digoxigenin (DIG), Helicobacter pylori, tHDA-ELISA, ureC, bacterial DNA, bacterial enzyme, digoxigenin, helicase, sulfonic acid derivative, amplicon, article, bacterium culture, bacterium detection, bacterium identification, biotinylation, colorimetry, controlled study, diagnostic accuracy, diagnostic approach route, diagnostic test, DNA hybridization, DNA probe, early diagnosis, enzyme denaturation, enzyme immobilization, enzyme linked immunosorbent assay, enzyme substrate, gene amplification, Helicobacter pylori, microtiter plate assay, nonhuman, nucleotide sequence, priority journal, stomach biopsy, thermophilic helicase dependent isothermal DNA amplification, Enzyme-Linked Immunosorbent Assay, Helicobacter Infections, Helicobacter pylori, Humans, Nucleic Acid Amplification Techniques, Sensitivity and Specificity, Urease
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 18 Apr 2015 03:46
Last Modified: 11 Feb 2017 08:18
URI: http://eprints.goums.ac.ir/id/eprint/2366

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