Multiplex PCR Assay for Rapid Detection and Genotyping of
            <i>Helicobacter pylori</i>
            Directly from Biopsy Specimens

Chattopadhyay, Santanu; Patra, Rajashree; Ramamurthy, T.; Chowdhury, Abhijit; Santra, Amal; Dhali, Gopal Krishna; Bhattacharya, Sujit; Berg, Douglas E.; Nair, G. Balakrish; Mukhopadhyay, Asish K.

doi:10.1128/jcm.42.6.2821-2824.2004

Cited by 100 publications

(76 citation statements)

References 13 publications

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“…Multiplex PCR reactions were performed with the use of specific primers for amplification of the cagA gene and subtypes of the vacA (s1/2 and m1/2) gene in biopsy specimens [18] . The tissue samples were suspended in lysis buffer (10 mmol/L Tris-HCl, 0.1 mol/L EDTA, 5% SDS, 100-200 μg/mL proteinase K).…”

Section: Multiplex Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Gastric ghrelin in relation to gender, stomach topographyand Helicobacter pylori in dyspeptic patients

Stec−Michalska¹,

Malicki²,

Michalski³

et al. 2009

WJG

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AIM:To investigate the level of gastric ghrelin in stomach mucosa of dyspeptic patients in relation to Helicobacter pylori (H pylori ) infection, bacterial cytotoxicity, topography and gender. METHODS:The study comprised 40 premenopausal women (19 H pylori positive) and 48 men (17 H pylori positive) with functional dyspepsia. All gastric biopsy specimens revealed normal mucosa or non-atrophic gastritis. Gastric ghrelin concentration was determined by Enzyme linked immunosorbent assay. The cagA and vacA strains of bacterial DNA were identified by multiplex polymerase chain reaction. RESULTS:In general, infection with H pylori caused an increase in gastric ghrelin level regardless of gender and stomach topography. Significantly more hormone was present in both, non-infected and H pylori positive female samples, as compared to males. The distribution of bacterial strains showed cagA (+) vacA s1m1 and cagA (-) vacA s2m2 genotypes as the most common infections in the studied population. A tendency to higher ghrelin levels was observed in less cytotoxic (cagA negative) strain-containing specimens from the antrum and corpus of both gender groups (without statistical significance). CONCLUSION: An increase in gastric ghrelin levelsat the stage of non-atrophic gastritis in H pylori positive patients, especially in those infected with cagA (-) strains, can exert a gastroprotective effect.

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Section: Multiplex Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Gastric ghrelin in relation to gender, stomach topographyand Helicobacter pylori in dyspeptic patients

Stec−Michalska¹,

Malicki²,

Michalski³

et al. 2009

WJG

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show abstract

“…For the genetic identification of H. pylori several PCR assay methods have been developed [5]. In a study vacA and cagA were investigated by multiplex PCR in directly from gastric biopsy specimens, for genotyping [6]. In another study H. pylori was detected by using one step multiplex PCR targeted to urease A gene, 16S ribosomal RNA and hpaA gene [7].…”

Section: Introductionmentioning

confidence: 99%

A Multiplex-Urease PCR Assay for Detection of Helicobacter pylori Infection Directly from Gastric Biopsy Specimens and Comparison of Multiplex- Urease PCR Results with Rapid Urease Test and Histopathology

Oktem-Okullu¹,

Tıftıkcı²,

Saruç³

et al. 2017

Clin Microbiol

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Purpose: To develop a multiplex-urease PCR assay specific for ureA and ureB genes directly from gastric biopsy specimens and compare the results of multiplex-urease PCR assay with rapid urease test (RUT) and histopathology.Methodology: This study was conducted on 109 patients. A RUT was run; histopathological staining and multiplex urease PCR were applied to biopsy specimen to detect H. pylori. Multiplex-urease PCR, RUT and histopathology results were compared by calculating Cohen's kappa coefficient.Results: A multiplex-urease PCR assay specific for ureA and ureB genes was developed to detect H. pylori directly from biopsy specimens. Cohen's kappa coefficient results of histopathological staining and multiplex-urease PCR indicate a substantial agreement. There was a moderate agreement between the results of histopathological staining and RUT results. There is a fair agreement between the multiplex urease PCR and the RUT results. Furthermore, the multiplex-urease PCR can detect H. pylori in some samples that are identified as negative by the rapid urease test and histopathological staining method. Moreover, in some patient samples ureA could not be detected while ureB detected. Conclusion:Multiplex-urease PCR assay was developed to detect ureA and ureB genes of H. pylori directly from gastric biopsy specimens. Comparison results indicated that detection rate of H. pylori with multiplex-urease PCR and histopathological staining is higher than the RUT. Moreover, it is critical to apply RUT test to biopsy specimens taken from both antrum and corpus part as in multiplex urease PCR assay. Furthermore, developing a multiplex PCR for both ureA and ureB is essential to detect active H. pylori infection.

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“…Based on the genetic characteristics and disease outcome, there are significant geographic differences among H. pylori strains. Indian H. pylori strains are genetically distinct from those from east Asia and the West (4,16). Several putative virulence-associated factors, including alleles in the cag pathogenicity island (PAI) of H. pylori, contribute to its pathogenesis and augment the risk for gastric adenocarcinoma (14).…”

mentioning

confidence: 99%

Antimicrobial Activity of Curcumin against Helicobacter pylori Isolates from India and during Infections in Mice

Kundu

Swarnakar

et al. 2009

Antimicrob Agents Chemother

Self Cite

388

203

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Treatment failure is a major cause of concern for the Helicobacter pylori-related gastroduodenal diseases like gastritis, peptic ulcer, and gastric cancer. Curcumin, diferuloylmethane from turmeric, has recently been shown to arrest H. pylori growth. The antibacterial activity of curcumin against 65 clinical isolates of H. pylori in vitro and during protection against H. pylori infection in vivo was examined. The MIC of curcumin ranges from 5 g/ml to 50 g/ml, showing its effectiveness in inhibiting H. pylori growth in vitro irrespective of the genetic makeup of the strains. The nucleotide sequences of the aroE genes, encoding shikimate dehydrogenase, against which curcumin seems to act as a noncompetitive inhibitor, from H. pylori strains presenting differential curcumin MICs showed that curcumin-mediated growth inhibition of Indian H. pylori strains may not be always dependent on the shikimate pathway. The antimicrobial effect of curcumin in H. pylori-infected C57BL/6 mice and its efficacy in reducing the gastric damage due to infection were examined histologically. Curcumin showed immense therapeutic potential against H. pylori infection as it was highly effective in eradication of H. pylori from infected mice as well as in restoration of H. pylori-induced gastric damage. This study provides novel insights into the therapeutic effect of curcumin against H. pylori infection, suggesting its potential as an alternative therapy, and opens the way for further studies on identification of novel antimicrobial targets of curcumin.

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Multiplex PCR Assay for Rapid Detection and Genotyping of Helicobacter pylori Directly from Biopsy Specimens

Cited by 100 publications

References 13 publications

Gastric ghrelin in relation to gender, stomach topographyand Helicobacter pylori in dyspeptic patients

Gastric ghrelin in relation to gender, stomach topographyand Helicobacter pylori in dyspeptic patients

A Multiplex-Urease PCR Assay for Detection of Helicobacter pylori Infection Directly from Gastric Biopsy Specimens and Comparison of Multiplex- Urease PCR Results with Rapid Urease Test and Histopathology

Antimicrobial Activity of Curcumin against Helicobacter pylori Isolates from India and during Infections in Mice

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