Comparison of three PCR methods for detection of<i>Helicobacter pylori</i>DNA and detection of<i>cagA</i>gene in gastric biopsy specimens

Smith, Stella I.; Oyedeji, Kolawole S.; Ao, Arigbabu; Cantet, Franck; Mégraud, Françis; Ojo, OO; Ao, Uwaifo; Otegbayo, JA; Ola, S O; Ao, Coker

doi:10.3748/wjg.v10.i13.1958

Cited by 67 publications

(48 citation statements)

References 7 publications

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“…We evaluated the 26 primer pairs previously reported for detection of H. pylori in clinical samples and included 2 primer pairs for the ureA gene, 2 for the 860-bp DNA gene, 3 for the16S rRNA gene, 1 for the 26K species-specific antigen gene, 8 for the vacA gene, 6 for the cagA gene, 3 for the glmM gene, and 1 for the adhesin gene. We used PCR conditions exactly matching those described by the authors reporting their use (7,10,16,20,26,27,29,32,34,40,43,47,48,55).…”

Section: Methodsmentioning

confidence: 99%

“…A number of target genes have been proposed as candidates for the PCR detection of H. pylori, including the 16S rRNA gene, the 26K species-specific antigen gene, the glmM gene, the ureA gene, the ureB gene, the vacA gene, and the cagA gene (see Table S1 in the supplemental material) (7,16,20,26,27,29,32,34,40,43,47,48,55). Although previous reports generally report good sensitivity and/or specificity of the primer pairs used, systematic studies comparing different PCR primer pairs are rare using very well-characterized cases (e.g., negative or positive by multiple tests) (12,45).…”

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confidence: 99%

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Unreliability of Results of PCR Detection of Helicobacter pylori in Clinical or Environmental Samples

Sugimoto

Abudayyeh

et al. 2009

J Clin Microbiol

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The aim of this study was to compare published Helicobacter pylori primer pairs for their ability to reliably detect H. pylori in gastric biopsy specimens and salivary samples. Detection limits of the 26 PCR primer pairs previously described for detection of H. pylori DNA in clinical samples were determined. Sensitivity and specificity were determined using primers with detection limits of <100 CFU/ml using 50 H. pylori-positive and -negative (by concordance by culture and histology) coded gastric biopsy specimens. These results were then confirmed with gastric biopsy specimens and saliva from patients with confirmed H. pylori status. Five of the twenty-six previously reported primer pairs (HP64-f/HP64-r, HP1/HP2, EHC-U/EHC-L, VAG-F/VAG-R, and ICT37/ICT38) had detection limits of <100 CFU/ml in the presence of gastric tissue. None had 100% specificity or sensitivity; all produced false-positive results. The HP64-f/HP64-r for ureA and HP1/HP2 for 16S rRNA individually had sensitivities and specificities of >90% with gastric biopsy specimens. No combinations of primer pairs improved the results. Using these five primer pairs, 54% of the positive saliva samples were determined to be false positive; both the HP64-f/HP64-r and the HP1/HP2 sets produced false positives with saliva. We conclude that clinicians should not rely on results using current PCR primers alone to decide the H. pylori status of an individual patient or as a basis for treatment decisions. The results of studies based on PCR identification of H. pylori in environmental samples should be viewed with caution. Possibly, specific primers sets can be identified based on the presence of multiple putative virulence factor genes.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Unreliability of Results of PCR Detection of Helicobacter pylori in Clinical or Environmental Samples

Sugimoto

Abudayyeh

et al. 2009

J Clin Microbiol

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“…La genotipifi cación del polimorfi smos IL-1B-511 se realizó a partir de ADN genómico extraí-do de las biopsias del antro gástrico 22 , por medio de discriminación alélica con PCR en tiempo real, y se emplearon las combinaciones de iniciadores y sondas reportadas por Johnson et al 23 . Cada reacción se realizó con un volumen fi nal de 20 µl con 40 ng de ADN genómico, 300 nM de cada iniciador, 250 nM de cada sonda, 10 La genotipifi cación del polimorfi smo antagonista del receptor de IL-1 (IL-1RN) se determinó mediante amplifi cación por PCR según la metodología descrita por Perri et al 24 , y se visualizaron mediante electroforesis en gel de agarosa al 25%.…”

Section: Análisis De Los Polimorfi Smos Il-1b-511 E Il1-rnunclassified

Asociación de los polimorfismos IL-1B-511 e IL-1RN y Helicobacter pylori CagA positivo con cáncer gástrico en una zona de riesgo alto en Colombia

et al. 2011

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“…The presence of H. pylori DNA in biopsy samples were detected by PCR amplification of H. pylori ureC gene (Table 1). In addition, cagA gene was detected by PCR amplification of a 297-bp region in the cagA gene as previously described (Table 1) (16,17). H. pylori ATCC49503 was used as positive control.…”

Section: Preparation Of Samples For Polymerase Chain Reactionmentioning

confidence: 99%

Helicobacter Pylori and CagA: Relationships With Esophageal and Gastroduodenal Disorders in Iranian Patients

Teymournejad¹,

Shokoohizadeh

Mobarez³

et al. 2015

Int J Enteric Pathog

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Background:The severity of Helicobacter pylori infection is associated with virulence factors of the bacteria and host immune response. H. pylori has several virulence factors which a number of them are essential to emerge clinical outcomes. Cytotoxin-associated gene A (CagA) is the most important H. pylori virulence factor. Objectives: The aim of our study was to assess a significant relationship between presence of cagA and severity of clinical manifestation in esophageal and gastroduodenal disorders. Patients and Methods: A total of 240 gastric biopsies were collected between March 2012 and August 2013 from Tehran's hospitals. Three sets of biopsy specimens were obtained from the antrum and rapid urease tests, histological examination, Polymerase Chain Reaction (PCR) assay were performed on the biopsy specimens. Results: One hundred and eight (45%) of biopsy specimens were positive with rapid urease test and ureC gene PCR. Moreover, thirty eight (35.1%) of positive specimens had cagA gene. The rate of gastric and duodenum inflammation was more in patients who carried CagA positive H. pylori strains. Whereas less inflammation and sever lesions in esophagus were found in CagA negative H. pylori strains. Conclusions: Our study demonstrates a strong relationship between CagA and esophageal and gastroduodenal disorders. The number of CagA negative H. pylori was larger than CagA positive in esophagus lesion grade A, C, and D. Therefore, cagA may have a protective effect on some esophageal diseases. In addition, the number of CagA positives was larger than CagA negative H. pylori in gastric antrum and duodenum ulcer. Thus, CagA play a role to emerge peptic and duodenal ulcers.

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Comparison of three PCR methods for detection ofHelicobacter pyloriDNA and detection ofcagAgene in gastric biopsy specimens

Abstract: This method is useful for correctly identifying infections caused by H pylori and can be easily applied in our laboratory for diagnostic purposes.

Cited by 67 publications

References 7 publications

Unreliability of Results of PCR Detection of Helicobacter pylori in Clinical or Environmental Samples

Unreliability of Results of PCR Detection of Helicobacter pylori in Clinical or Environmental Samples

Asociación de los polimorfismos IL-1B-511 e IL-1RN y Helicobacter pylori CagA positivo con cáncer gástrico en una zona de riesgo alto en Colombia

Helicobacter Pylori and CagA: Relationships With Esophageal and Gastroduodenal Disorders in Iranian Patients

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