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      An improved quantitative real-time polymerase chain reaction technology for Helicobacter pylori detection in stomach tissue and its application value in clinical precision testing

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          Abstract

          Background

          Helicobacter pylori ( H. pylori) infection is a serious human health threat. The empiric H. pylori treatment paradigm guided by traditional testing technologies has led to antibiotic resistance. Here, we improved the qPCR method to provide technical support for precision H. pylori diagnosis and treatment.

          Methods

          Two pairs of primers and probes targeting the glmM gene were designed to detect H. pylori, and a multiplex qPCR method was established for virulence factor detection. Then, a rapid urease test (RUT), culturing and qPCR were performed on 141 specimens collected from Xinqiao Hospital of China in 2017 to evaluate the qPCR detection capability. Finally, the H. pylori infectious amount and virulence genes were detected by qPCR.

          Results

          1. The improved qPCR method which used two pairs of primers had a higher detection rate (100%) and better accuracy ( p = 0.000), compared with the qPCR using a pair of primers. It also had better consistency with the bacterial culture than with RUT (Kappa =0.440, p < 0.001). 2. The H. pylori infectious amount was significantly positively associated with gastritis in corpus ( p = 0.003) and gastric erosion ( p = 0.043). The H. pylori infectious amount in gastric precancerous patients was significantly lower than that in H. pylori-positive patients ( p < 0.05), and the infectious H. pylori-vacA s1+ amount was significantly greater than that of H. pylori-vacA s1- (p < 0.05). 3. The vacA s1 frequency was significantly higher than that of vacA m1/cagA+/babA2+ in chronic superficial gastritis ( p = 0.000), peptic ulcer ( p = 0.037) and gastric erosion ( p = 0.009). The H. pylori-vacA+/cagA+/babA2+ frequency showed a significant positive correlation ( p < 0.05).

          Conclusions

          The H. pylori infectious amount and presence of H. pylori virulence factors showed complex correlations with gastric disease occurrence and development. The improved qPCR with good detection performance can be used for quantitative H. pylori detection and testing for the virulence genes vacA s1, vacA m1, cagA and babA2 simultaneously. These findings will provide valuable information for disease diagnosis and treatment.

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          Most cited references33

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          Oncogenic mechanisms of the Helicobacter pylori CagA protein.

          Infection with strains of Helicobacter pylori that carry the cytotoxin-associated antigen A (cagA) gene is associated with gastric carcinoma. Recent studies have shed light on the mechanism through which the cagA gene product, CagA, elicits pathophysiological actions. CagA is delivered into gastric epithelial cells by the bacterial type IV secretion system, where it deregulates the SHP2 oncoprotein. Intriguingly, CagA is noted for its variation, particularly at the SHP2-binding site, which could affect the potential of different strains of H. pylori to promote gastric carcinogenesis.
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            Fifth Chinese National Consensus Report on the management of Helicobacter pylori infection.

            Since the 'Fourth Chinese National Consensus Report on the management of H. pylori infection' was published in 2012, three important consensuses (Kyoto global consensus report on H. pylori gastritis, The Toronto Consensus for the Treatment of H. pylori Infection in Adults and Management of H. pylori infection-the Maastricht V/Florence Consensus Report) have been published regarding the management of H. pylori infection.
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              Diagnosis of Helicobacter pylori infection: Current options and developments.

              Accurate diagnosis of Helicobacter pylori (H. pylori) infection is a crucial part in the effective management of many gastroduodenal diseases. Several invasive and non-invasive diagnostic tests are available for the detection of H. pylori and each test has its usefulness and limitations in different clinical situations. Although none can be considered as a single gold standard in clinical practice, several techniques have been developed to give the more reliable results. Invasive tests are performed via endoscopic biopsy specimens and these tests include histology, culture, rapid urease test as well as molecular methods. Developments of endoscopic equipment also contribute to the real-time diagnosis of H. pylori during endoscopy. Urea breathing test and stool antigen test are most widely used non-invasive tests, whereas serology is useful in screening and epidemiological studies. Molecular methods have been used in variable specimens other than gastric mucosa. More than detection of H. pylori infection, several tests are introduced into the evaluation of virulence factors and antibiotic sensitivity of H. pylori, as well as screening precancerous lesions and gastric cancer. The aim of this article is to review the current options and novel developments of diagnostic tests and their applications in different clinical conditions or for specific purposes.
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                Author and article information

                Contributors
                pettyfeng@hotmail.com
                Journal
                BMC Biotechnol
                BMC Biotechnol
                BMC Biotechnology
                BioMed Central (London )
                1472-6750
                22 June 2020
                22 June 2020
                2020
                : 20
                : 33
                Affiliations
                [1 ]GRID grid.410570.7, ISNI 0000 0004 1760 6682, Department of Clinical Microbiology and Immunology, Faculty of Pharmacy and Medical Laboratory Sciences, , Third Military Medical University (Army Medical University), ; No. 30 Gaotanyan Street, Chongqing, 400038 People’s Republic of China
                [2 ]GRID grid.203458.8, ISNI 0000 0000 8653 0555, Department of Digestive Disease Center, , The Third Affiliated Hospital of Chongqing Medical University (General Hospital), ; Chongqing, China
                Author information
                https://orcid.org/0000-0003-2451-6936
                Article
                624
                10.1186/s12896-020-00624-z
                7310109
                32571272
                2d9af072-8e40-4160-9a5b-a379eb33fb9f
                © The Author(s) 2020

                Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

                History
                : 8 December 2019
                : 19 May 2020
                Funding
                Funded by: National Natural Science Foundation of China (CN)
                Award ID: 81601832
                Award Recipient :
                Categories
                Methodology Article
                Custom metadata
                © The Author(s) 2020

                Biotechnology
                quantitative detection,precision testing,helicobacter pylori
                Biotechnology
                quantitative detection, precision testing, helicobacter pylori

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