Genotyping and Association of the Presence of Helicobacter Pylori in Dental Plaque and Gastric Biopsy Specimens in Dyspeptic Patients by PCR-RFLP Technique in Southwest of Iran

Abstract The oral cavity can act as an extra gastric reservoir for H pylori, and also the presence of the bacteria in the oral cavity is associated with a higher risk of dental caries development. The aim of this study was to determine the genotype and evaluate the association of the presence of H. pylori in dental plaque and gastric biopsy specimens in dyspeptic patients in Ahvaz, Southwest of Iran. In this study, 106 patients with recruited dyspeptic complaints were selected and from each patient, two gastric antral biopsy specimens and two dental plagues were examined. The presence of H. pylori was identified by the Rapid Urease Test (RUT) and the amplification of ureAB and 16S rRNA genes. Also, to verify a hypothetical mouth-to-stomach infection route, the enzymatic digestions of three genes of cagA, vacA, and ureAB in H. pylori strains isolated from dental plaques and stomach samples were compared for each same case. H. pylori was found in the stomach of 52.8% (56 /106) and the dental plaques of 17.9% (19/106) of the studied cases. On the other hand, H. pylori was recognized in the stomach of all 19 cases with oral colonization. Following a combination of restriction fragment length polymorphism (RFLP) patterns of these three known genes on stomach and dental plague samples, 14 and 11 unique patterns were seen, respectively. However, for all H. pylori-positive cases (19), the comparison of RLFP patterns of these genes in the dental plaque and gastric biopsy specimens was different for the same case. This study showed, no significant association was observed between the presence of H. pylori in dental plaque and the stomach of the same case.

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Mutational Analysis of Clarithromycin and Levofloxacin Resistance in Helicobacter pylori from Gastric Biopsy Specimens in a Tertiary Care Hospital in Dhaka, Bangladesh

Bangladesh Journal of Medical Microbiology ◽

10.3329/bjmm.v13i1.51780 ◽

2019 ◽

Vol 13 (1) ◽

pp. 12-19

Author(s):

Nusrat Noor Tanni ◽

Shaheda Anwar ◽

Sharmeen Ahmed ◽

Kakali Halder ◽

Maherun Nesa ◽

...

Keyword(s):

Gene Mutations ◽

Gastric Biopsy ◽

23S Rrna ◽

Rapid Urease Test ◽

Care Hospital ◽

Gyra Gene ◽

Biopsy Specimens ◽

Urease Test ◽

Gastric Biopsies ◽

H Pylori

Background: Clarithromycin and Levofloxacin are most frequently included in the standard triple therapies for H. pylori eradication in our country. Resistance to clarithromycin and fluoroquinolones are particularly related with treatment failure. Objectives: The objective of this study was to detect, clarithromycin and levofloxacin resistance associated with gene mutations in H. pylori directly from gastric biopsies using an allele specific primer-PCR (ASP-PCR) assay. Materials and Methods: Gastric biopsy specimens were collected from 143 adult dyspeptic patients, from Department of Gastroenterology, BSMMU and Dhaka Medical College Hospital (DMCH), during the period of March, 2018 to February, 2019. H. pylori was identified by rapid urease test, ureC gene by PCR, histological staining and culture. ASP-PCR was used to identify 23S rRNA gene and gyrA gene mutation predictive of clarithromycin and levofloxacin resistant H. pylori respectively. Results: H. pylori positive cases were 32.9% based on the case definition used in the study. Among 42 ureC positive H. pylori cases, point mutations in 23Sr RNA gene for clarithromycin resistance were detected only at A2142G position in 9 (21.4%) cases and gyrA gene mutations for levofloxacin resistance were detected in 16 (38.1%) cases. Only 1 (2.4%) case had mutation both in 23Sr RNA and gyrA gene. Conclusion: Those findings may guide toward the therapeutic choices in our country. PCR based diagnostic assays can be the alternative approach for rapid detection of antibiotic resistances of H. pylori directly from gastric biopsies, where culture and susceptibility tests are not routinely performed. Bangladesh J Med Microbiol 2019; 13 (1): 12-19

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Helicobacter pyloridetection and genotyping in gastric biopsy specimens from Chennai patients (India)

Canadian Journal of Microbiology ◽

10.1139/w08-108 ◽

2009 ◽

Vol 55 (2) ◽

pp. 126-132 ◽

Cited By ~ 3

Author(s):

Gopal Udhayakumar ◽

Cinghu Senthilkumar ◽

Venkatraman Jayanthi ◽

Niranjali Devaraj ◽

Halagowder Devaraj

Keyword(s):

Biopsy Specimen ◽

South India ◽

Gastric Biopsy ◽

Rapid Urease Test ◽

Caga Gene ◽

Potential Value ◽

Biopsy Specimens ◽

Urease Test ◽

H Pylori ◽

Subtype A

The aim of the present study was to investigate the prevalence of Helicobacter pylori infection and the correlation between cagA and vacA (mid-region) genotypes with different clinical outcomes from Chennai, India, patients. Biopsies from the antrum were taken to assess the current H. pylori status by histology, rapid urease test (RUT), and PCR. The RUT and PCR analyses were carried out on a single biopsy specimen. Fasting sera were obtained from all patients and H. pylori status was determined by using ELISA. In addition, the correlations between cagA and vacA genotypes and the consequence of H. pylori infection were statistically examined. Prevalence of the cagA gene was found in 96% (90/94) of patients, and the vacA m2 subtype occurred in 60% (56/94), whereas 32% (30/94) showed the vacA m1 subtype. A significant association between the cagA and vacA m2 region (χ2 = 5.556; p < 0.01) was found in ulcer patients. The vacA m2 genotype showed a near-significant value (χ2 = 3.943; p < 0.047) in ulcer patients when compared with vacA m1. These findings suggest that H. pylori strains with the vacA m2 region were predominant in South India, especially in and around Chennai. This study also showed that PCR has a potential value for studying the cagA gene directly from biopsy specimens.

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An evaluation of the usefulness of invasive and non-invasive methods used to diagnose Helicobacter spp. infections in dogs

Polish Journal of Veterinary Sciences ◽

10.1515/pjvs-2017-0059 ◽

2017 ◽

Vol 20 (3) ◽

pp. 491-499 ◽

Cited By ~ 2

Author(s):

M. Jankowski ◽

J. Spużak ◽

K. Kubiak ◽

K. Glińska-Suchocka ◽

M. Biernat

Keyword(s):

Nested Pcr ◽

Gastric Biopsy ◽

Diagnostic Methods ◽

Rapid Urease Test ◽

Pcr Assay ◽

Non Invasive ◽

Biopsy Specimens ◽

Urease Test ◽

Invasive Method ◽

Pcr Method

AbstractThe aim of this study was to assess the suitability of invasive and non-invasive methods used to diagnose Helicobacter spp. in the stomachs of dogs. The study was carried out on 30 dogs of both sexes and different breeds, between one and 15 years old. A histopathologic examination, a microbiological culture, a rapid urease test, a direct bacteriological preparation and a nested PCR assay were carried out. Gastric Helicobacter spp. was identified in gastric biopsy specimens from 16 (53.3%) dogs using direct bacteriological preparation, in four (13.3%) dogs based on a culture, in 23 (76.6%) dogs using the rapid urease test and in 21 (70,0%) dogs based on a histopathological assessment of the biopsy specimens. The nested PCR of the gastric biopsy specimens revealed gastric Helicobacter spp. in all the dogs (100%). A saliva PCR assay revealed gastric Helicobacter spp. in 23 (76.6%) dogs, while stool PCR revealed the bacterium in seven (23.3%) dogs. We found that invasive methods were more accurate than non-invasive methods in detecting a Helicobacter spp. infection in dogs. In addition, the nested PCR method used to evaluate the gastric mucosal biopsy specimens was the most accurate test for detecting Helicobacter spp. It was further found that the PCR-based saliva assay was the best non-invasive method for detecting Helicobacter spp. However, taking into consideration that most of the diagnostic methods used to detect this bacterium have drawbacks, at least two diagnostic methods should be used to detect Helicobacter spp. as is done in human medicine.

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Polymerase chain reaction assay for the detection of Helicobacter pylori in gastric biopsy specimens: comparison with culture, rapid urease test, and histopathological tests.

Gut ◽

10.1136/gut.35.7.905 ◽

1994 ◽

Vol 35 (7) ◽

pp. 905-908 ◽

Cited By ~ 56

Author(s):

R Fabre ◽

I Sobhani ◽

P Laurent-Puig ◽

N Hedef ◽

N Yazigi ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Helicobacter Pylori ◽

Polymerase Chain Reaction Assay ◽

Gastric Biopsy ◽

Rapid Urease Test ◽

Chain Reaction ◽

Biopsy Specimens ◽

Urease Test ◽

Polymerase Chain

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Helicobacter pyloriIs Not Eradicated after Triple Therapy: A Nested PCR Based Study

BioMed Research International ◽

10.1155/2014/483136 ◽

2014 ◽

Vol 2014 ◽

pp. 1-8 ◽

Cited By ~ 10

Author(s):

Saurabh Kumar Patel ◽

Girish Narayan Mishra ◽

Chandra Bhan Pratap ◽

Ashok Kumar Jain ◽

Gopal Nath

Keyword(s):

Triple Therapy ◽

Nested Pcr ◽

Restriction Pattern ◽

Rapid Urease Test ◽

Serological Tests ◽

Biopsy Specimens ◽

Urease Test ◽

Before And After ◽

H Pylori ◽

Antral Biopsy

Detection ofHelicobacter pyloriafter triple therapy is usually carried out by either rapid urease test (RUT), urea breath test (UBT), histology, bacterial isolation, and single round PCR or serological tests. In this study, antral biopsy specimens from 25 patients were tested forH. pyloriby RUT, culture, histology, and nested PCR in their antral biopsy specimens before and after treatment. Three genes, namely, heat shock protein (hsp60), phosphoglucosamine mutase (ureC), and flagellar export ATP synthase (fliI) ofH. pyloriwere targeted. Of the 25 antral biopsy specimens, the RUT, culture, histology, and nested PCR positivity dropped from 81.8% to 12%, 31% to 0%, 100 to 84%, and 100% to 92%, respectively, before and after therapy. Further,hsp60specific amplicons from 23 out of 25 patients gave identical restriction pattern, while 6fliIand 1ureCspecific amplicon produced different restriction pattern. Furthermore, variations infliIgene sequences inH. pyloriafter treatment were also confirmed by sequencing and comparedin silico. Nested PCR based detection ofH. pyloriis more sensitive method to detectH. pyloriafter therapy than culture, RUT, and histology. Further, this study suggests thatH. pyloriis not eradicated completely after triple therapy.

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Comparative study of rapid urease test and histopathological examination for detection of H. pylori infection

International Surgery Journal ◽

10.18203/2349-2902.isj20175413 ◽

2017 ◽

Vol 4 (12) ◽

pp. 4071

Author(s):

Virendra S. Athavale ◽

Vinit Singh ◽

Shivmurti N. Khandalkar ◽

Dakshayani S. Nirhale ◽

Aditya Lad ◽

...

Keyword(s):

Histopathological Examination ◽

Good Alternative ◽

Gastric Biopsy ◽

Rapid Urease Test ◽

Study Group ◽

Medical College ◽

Urease Test ◽

Resource Poor ◽

H Pylori

Background: Aims and objectives of the study was to study the incidence of H. pylori infection in our Hospital. To find out the sensitivity and specificity of rapid urease test (RUT) and histopathological examination for the detection of H. pylori on gastric biopsy. To study the effect of anti H. pylori drugs by performing follow-up endoscopy in terms of positive or negative Rapid Urease Test and Histopathological Examination report.Methods: The study was conducted at Dr. D. Y. Patil Medical College and Hospital, DPU University, for a period of 2 years (from July 2012-September 2014) and is a prospective and comparative randomized type of study using 100 patients. The study was approved by the Institute’s Ethics Committee.Results: Data analysis showed that: In our study 84 patients (84%) were detected positive by RUT method. In our study 83 patients (83 %) were detected positive by HPE method. There was an association between RUT and HPE finding in study group with the sensitivity being 96.38 % and specificity being 74.47 %, PPV was 95.24% and NPV was 81.25%. At the time of follow up after taking anti H pylori treatment, 79 patients were RUT negative and 8 were RUT positive who were defaulters (not taken complete dose) and again started on treatment with the accuracy of the drug being 98.85 %. At the time of follow up after taking anti H pylori treatment 78 patients were HPE negative and 9 were HPE positive who were defaulters (not taken complete dose) and again started on treatment. There was an association between RUT and HPE finding at follow up with the sensitivity being 88.89 % and specificity being 100%, PPV was 100% and NPV was 98.73%.Conclusions: Our study reveals that RUT is accurate for the diagnosis of H. pylori infection and its use will serve as a good alternative to histology in management of patients with dyspepsia in resource poor environments, except in patients who need histology for reasons other than H. pylori diagnosis.

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