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Cited by 44Helicobacter pylori-related gastroenterology studiesMolecular Biology Techniques and ApplicationsBacterial Identification and Susceptibility Testing

Specific and Sensitive Detection of H. pylori in Biological Specimens by Real-Time RT-PCR and In Situ Hybridization

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Liu, Hui, Rahman, Arifur, Semino-Mora, Cristina, Doi, Sonia Q., Dubois, Andre·2008·PLoS ONE
DOI 10.1371/journal.pone.0002689PMC PMC2481290
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Abstract

PCR detection of H. pylori in biological specimens is rendered difficult by the extensive polymorphism of H. pylori genes and the suppressed expression of some genes in many strains. The goal of the present study was to (1) define a domain of the 16S rRNA sequence that is both highly conserved among H. pylori strains and also specific to the species, and (2) to develop and validate specific and sensitive molecular methods for the detection of H. pylori. We used a combination of in silico and molecular approaches to achieve sensitive and specific detection of H. pylori in biologic media. We sequenced two isolates from patients living in different continents and demonstrated that a 546-bp domain of the H. pylori 16S rRNA sequence was conserved in those strains and in published sequences. Within this conserved sequence, we defined a 229-bp domain that is 100% homologous in most H. pylori strains available in GenBank and also is specific for H. pylori. This sub-domain was then used to design (1) a set of high quality RT-PCR primers and probe that encompassed a 76-bp sequence and included at least two mismatches with other Helicobacter sp. 16S rRNA; and (2) in situ hybridization antisense probes. The sensitivity and specificity of the approaches were then demonstrated by using gastric biopsy specimens from patients and rhesus monkeys. This H. pylori-specific region of the 16S rRNA sequence is highly conserved among most H. pylori strains and allows specific detection, identification, and quantification of this bacterium in biological specimens.

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  • Studies investigating human brain tissue confirm that both hypo- and hyper-methylation of DNA are associated with AD.

    “Studies investigating human brain tissue confirm that both hypo– and hyper-methylation of DNA are associated with AD17,18.”

    From Integrated analysis of behavioral, epigenetic, and gut microbiome analyses in AppNL-G-F, AppNL-F, and wild type mice (2021) · cites this paper as [18]

    The Introduction and Discussion address detection of H. pylori, while the Results section on DNA fingerprinting concerns bacterial strains. Searches for DNA methylation, Alzheimer’s disease, and human brain tissue found no passage addressing the claimed association; our copy therefore cannot verify it. (LLM, conf=0.78)

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References 38 · Cited by 44

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Authors
Liu, H., Rahman, A., Semino-Mora, C., Doi, S. Q., & Dubois, A.
Affiliations
Uniformed Services University of the Health Sciences (US)
Corresponding author
  • Liu, Hui
  • Rahman, Arifur
  • Doi, Sonia Q.
  • Dubois, Andre
ORCID
Liu, Hui
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2008-07-16
Volume
3
Issue
7
Pages
e2689
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0002689
PMC
PMC2481290
OpenAlex
W2160431243
Cited by
44
References
38
Citations by year
2023: 1 · 2022: 1 · 2021: 3 · 2020: 3 · 2019: 2 · 2018: 2 · 2017: 1 · 2016: 7 · 2015: 1 · 2014: 3 · 2013: 4 · 2012: 5
FWCI
3.87
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, public-domain)
  • PubMed Central (publishedVersion, public-domain)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • U.S. Department of Defense
  • National Institutes of Health
  • Uniformed Services University of the Health Sciences
  • National Cancer Institute (R01CA082312)
Topics
  • Health Sciences › Medicine › Surgery › Helicobacter pylori-related gastroenterology studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Bacterial Identification and Susceptibility Testing
Keywords
Helicobacter pylori, 16S rRNA, real-time RT-PCR, in situ hybridization, antisense probes, primer and probe design, gastric biopsy, rhesus monkeys, bacterial detection, genetic polymorphism
MeSH terms
Animals, Base Sequence, Biopsy, Cloning, Molecular, Gastritis, Macaca mulatta, Molecular Sequence Data, RNA, Ribosomal, 16S, Stomach, Helicobacter pylori, Helicobacter Infections, Conserved Sequence, In Situ Hybridization, Sequence Analysis, DNA, DNA Primers

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