Prevalence of the Helicobacter pylori babA2 Gene in Children Mainly Depends on the PCR Primer Set Used

Various polymerase chain reaction- (PCR-) based methods with varying positivity rates were designed to detect the Helicobacter pylori babA2 gene. To compare different primer sets, babA2 prevalence was determined in 279 H. pylori-positive pediatric samples using the 832 bp, 139 bp, and 271 bp PCR pri...

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Main Authors: Anja Šterbenc, Maja M. Lunar, Matjaž Homan, Boštjan Luzar, Nina Zidar, Mario Poljak
Format: Article
Language:English
Published: Hindawi Limited 2020-01-01
Series:Canadian Journal of Infectious Diseases and Medical Microbiology
Online Access:http://dx.doi.org/10.1155/2020/4080248
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spelling doaj-373c75a0abed4b83b5cd07ec123c9c372021-07-02T17:12:47ZengHindawi LimitedCanadian Journal of Infectious Diseases and Medical Microbiology1712-95321918-14932020-01-01202010.1155/2020/40802484080248Prevalence of the Helicobacter pylori babA2 Gene in Children Mainly Depends on the PCR Primer Set UsedAnja Šterbenc0Maja M. Lunar1Matjaž Homan2Boštjan Luzar3Nina Zidar4Mario Poljak5Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloška 4, Ljubljana 1000, SloveniaInstitute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloška 4, Ljubljana 1000, SloveniaDepartment of Gastroenterology, Hepatology and Nutrition, University Children’s Hospital, Faculty of Medicine, University of Ljubljana, Vrazov Trg 2, Ljubljana 1000, SloveniaInstitute of Pathology, Faculty of Medicine, University of Ljubljana, Korytkova 2, Ljubljana 1000, SloveniaInstitute of Pathology, Faculty of Medicine, University of Ljubljana, Korytkova 2, Ljubljana 1000, SloveniaInstitute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloška 4, Ljubljana 1000, SloveniaVarious polymerase chain reaction- (PCR-) based methods with varying positivity rates were designed to detect the Helicobacter pylori babA2 gene. To compare different primer sets, babA2 prevalence was determined in 279 H. pylori-positive pediatric samples using the 832 bp, 139 bp, and 271 bp PCR primer sets, resulting in 34.0%, 51.3%, and 79.6% prevalence of the babA2 gene, respectively. The babA2 status determined using the 832 bp and 139 bp PCR primer sets significantly correlated with bacterial density and activity of inflammation, whereas no such correlations were found using the 271 bp PCR primer set. The 139 and 832 bp PCR primer sets concordantly detected the babA2 gene in 93 cases; however, in comparison to the 832 bp PCR primer set, the 139 bp PCR primer set detected additional 50 babA2 cases, whereas only two 832 bp positive cases were missed. The 271 bp PCR primer set missed 32 babA2 cases that were 832 bp and/or 139 bp PCR positive, but tested solely positive in 109 cases. Interestingly, cloning of a subset of 271 bp PCR positive samples revealed amplification of the babA/B gene chimera. Hence, in our opinion, the 271 bp PCR protocol is not a reliable diagnostic tool for detecting the babA2 gene in children. Our results reaffirm previous observations that the use of certain babA2 PCR primer sets can significantly impact estimation of the prevalence and clinical relevance of the H. pylori babA2 gene in children, suggesting babA2 detection methods should be carefully selected.http://dx.doi.org/10.1155/2020/4080248
collection DOAJ
language English
format Article
sources DOAJ
author Anja Šterbenc
Maja M. Lunar
Matjaž Homan
Boštjan Luzar
Nina Zidar
Mario Poljak
spellingShingle Anja Šterbenc
Maja M. Lunar
Matjaž Homan
Boštjan Luzar
Nina Zidar
Mario Poljak
Prevalence of the Helicobacter pylori babA2 Gene in Children Mainly Depends on the PCR Primer Set Used
Canadian Journal of Infectious Diseases and Medical Microbiology
author_facet Anja Šterbenc
Maja M. Lunar
Matjaž Homan
Boštjan Luzar
Nina Zidar
Mario Poljak
author_sort Anja Šterbenc
title Prevalence of the Helicobacter pylori babA2 Gene in Children Mainly Depends on the PCR Primer Set Used
title_short Prevalence of the Helicobacter pylori babA2 Gene in Children Mainly Depends on the PCR Primer Set Used
title_full Prevalence of the Helicobacter pylori babA2 Gene in Children Mainly Depends on the PCR Primer Set Used
title_fullStr Prevalence of the Helicobacter pylori babA2 Gene in Children Mainly Depends on the PCR Primer Set Used
title_full_unstemmed Prevalence of the Helicobacter pylori babA2 Gene in Children Mainly Depends on the PCR Primer Set Used
title_sort prevalence of the helicobacter pylori baba2 gene in children mainly depends on the pcr primer set used
publisher Hindawi Limited
series Canadian Journal of Infectious Diseases and Medical Microbiology
issn 1712-9532
1918-1493
publishDate 2020-01-01
description Various polymerase chain reaction- (PCR-) based methods with varying positivity rates were designed to detect the Helicobacter pylori babA2 gene. To compare different primer sets, babA2 prevalence was determined in 279 H. pylori-positive pediatric samples using the 832 bp, 139 bp, and 271 bp PCR primer sets, resulting in 34.0%, 51.3%, and 79.6% prevalence of the babA2 gene, respectively. The babA2 status determined using the 832 bp and 139 bp PCR primer sets significantly correlated with bacterial density and activity of inflammation, whereas no such correlations were found using the 271 bp PCR primer set. The 139 and 832 bp PCR primer sets concordantly detected the babA2 gene in 93 cases; however, in comparison to the 832 bp PCR primer set, the 139 bp PCR primer set detected additional 50 babA2 cases, whereas only two 832 bp positive cases were missed. The 271 bp PCR primer set missed 32 babA2 cases that were 832 bp and/or 139 bp PCR positive, but tested solely positive in 109 cases. Interestingly, cloning of a subset of 271 bp PCR positive samples revealed amplification of the babA/B gene chimera. Hence, in our opinion, the 271 bp PCR protocol is not a reliable diagnostic tool for detecting the babA2 gene in children. Our results reaffirm previous observations that the use of certain babA2 PCR primer sets can significantly impact estimation of the prevalence and clinical relevance of the H. pylori babA2 gene in children, suggesting babA2 detection methods should be carefully selected.
url http://dx.doi.org/10.1155/2020/4080248
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