A Ajayi1, T Jolaiya2, S I Smith3,4. 1. Department of Molecular Biology and Biotechnology, Nigerian Institute of Medical Research, Yaba, Lagos, Nigeria. 2. Department of Microbiology, University of Lagos, Lagos, Nigeria. 3. Department of Molecular Biology and Biotechnology, Nigerian Institute of Medical Research, Yaba, Lagos, Nigeria. stellaismith@yahoo.com. 4. Mountain Top University, Makogi Oba, Ogun, Nigeria. stellaismith@yahoo.com.
Abstract
OBJECTIVE: Prompt diagnosis of Helicobacter pylori infection is essential for proper treatment and eradication of the pathogen because prolonged infection could lead to gastric cancer. Sensitive and cost effective diagnostic methods are key to guiding treatment options that will reduce mortality. This study was aimed at detecting H. pylori from biopsies of peptic ulcer patients. Real-time PCR using TaqMan and EvaGreen assays targeting 16S rRNA and ureA genes were used to detect H. pylori DNA extracted from 40 biopsy samples comprising 20 biopsies obtained from the antrum and 20 from the corpus of 20 patients undergoing endoscopy for duodenal ulcer investigation in Lagos, Nigeria. RESULTS: H. pylori was detected in 80% of the biopsy samples by combined cycle threshold (Ct) and melting temperature (Tm) values. Mean Ct value for ureA gene ranged from 21.40 to 37.53 and 22.71 to 35.44 for 16SrRNA gene. Average melting temperatures (Tm) of 81.57 and 82.90 °C among amplicons of ureA and 16S rRNA were observed respectively. H. pylori DNA was generally detected in biopsies collected from antrum and corpus. Real-time PCR in the diagnosis of H. pylori can be considered a simple, low cost and efficient alternative or addition to the gold standard.
OBJECTIVE: Prompt diagnosis of Helicobacter pylori infection is essential for proper treatment and eradication of the pathogen because prolonged infection could lead to gastric cancer. Sensitive and cost effective diagnostic methods are key to guiding treatment options that will reduce mortality. This study was aimed at detecting H. pylori from biopsies of peptic ulcerpatients. Real-time PCR using TaqMan and EvaGreen assays targeting 16S rRNA and ureA genes were used to detect H. pylori DNA extracted from 40 biopsy samples comprising 20 biopsies obtained from the antrum and 20 from the corpus of 20 patients undergoing endoscopy for duodenal ulcer investigation in Lagos, Nigeria. RESULTS:H. pylori was detected in 80% of the biopsy samples by combined cycle threshold (Ct) and melting temperature (Tm) values. Mean Ct value for ureA gene ranged from 21.40 to 37.53 and 22.71 to 35.44 for 16SrRNA gene. Average melting temperatures (Tm) of 81.57 and 82.90 °C among amplicons of ureA and 16S rRNA were observed respectively. H. pylori DNA was generally detected in biopsies collected from antrum and corpus. Real-time PCR in the diagnosis of H. pylori can be considered a simple, low cost and efficient alternative or addition to the gold standard.
Authors: Stella I Smith; Muinah A Fowora; Jesse A Otegbayo; Fatimah B Abdulkareem; Emmanuel A Omonigbehin; Akere Adegboyega; Monica Contreras; Rainer Haas Journal: Int J Mol Epidemiol Genet Date: 2011-01-15
Authors: Bea Van den Poel; Sarah Gils; Isabel Micalessi; Saskia Carton; Paul Christiaens; Pieter-Jan Cuyle; Veerle Moons; Gust Van Olmen; Annick Smismans; Claire Bourgain; Peter Bossuyt; Johan Frans Journal: Acta Clin Belg Date: 2019-10-29 Impact factor: 1.264