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

2019 ◽  
Vol 13 (1) ◽  
pp. 12-19
Author(s):  
Nusrat Noor Tanni ◽  
Shaheda Anwar ◽  
Sharmeen Ahmed ◽  
Kakali Halder ◽  
Maherun Nesa ◽  
...  

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

2021 ◽  
Author(s):  
Mojtaba Moosavian ◽  
Elyas Kushki ◽  
Tahereh Navidifar ◽  
Eskandar Hajiani ◽  
Mahdi Mandegari

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.


2009 ◽  
Vol 55 (2) ◽  
pp. 126-132 ◽  
Author(s):  
Gopal Udhayakumar ◽  
Cinghu Senthilkumar ◽  
Venkatraman Jayanthi ◽  
Niranjali Devaraj ◽  
Halagowder Devaraj

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.


2020 ◽  
Vol 29 (3) ◽  
pp. 59-64
Author(s):  
Hanaa M. El Maghraby ◽  
Samar Mohaseb

Background: Metronidazole is one of the antimicrobial drugs that can be used in combination with other drugs for eradication of Helicobacter pylori (H. pylori).Unfortunately, metronidazole resistance in H. plori is an increasing health problem which may be attributed to inactivation of many genes as rdx A gene. Objective: To determine the frequency of rdx A deletion mutation in H. pylori detected in infected patients attending at the Gastroenterology Unit, Zagazig University Hospitals. Methodology: Two gastric biopsies were taken from each enrolled patient by endoscopy. H.pylori detection was done by rapid urease test and polymerase chain reaction (PCR) amplification of 16S rRNA gene. Deletion mutation in rdx A gene was detected by conventional PCR. Results: Out of 134 doubled gastric biopsies obtained from 134 patients, 52.2% were positive for H. pylori. Epigastric pain, vomiting and gastritis were significantly associated with detection of H. pylori infection (p˂ 0.05). Deletion mutation of rdx A gene was detected in 28.6% of H. pylori positive specimens obtained from infected patients. Conclusion: Deletion mutation of rdx A gene is a frequent determinant of rdx A inactivation conferring metronidazole resistance among H. pylori.


2019 ◽  
Vol 8 (7) ◽  
pp. 1071 ◽  
Author(s):  
Izabela Korona-Glowniak ◽  
Halina Cichoz-Lach ◽  
Radoslaw Siwiec ◽  
Sylwia Andrzejczuk ◽  
Andrzej Glowniak ◽  
...  

The aim of this study was to investigate genetic diversity of Helicobacter pylori virulence markers to predict clinical outcome as well as to determine an antibiotic susceptibility of H. pylori strains in Poland. Gastric biopsies from 132 patients with gastrointestinal disorders were tested for presence of H. pylori with the use of rapid urease test, microbial culture, and polymerase chain reaction (PCR) detection. The genetic diversity of 62 H. pylori positive samples was evaluated by detection of cagA and PCR-typing of vacA and iceA virulence-associated genes. Most common H. pylori genotypes were cagA(+)vacAs1m2 (27.4%) and cagA(−)vacAs2m2 (24.2%). In logistic regression analysis, we recognized the subsequent significant associations: gastritis with ureC, i.e., H. pylori infection (p = 0.006), BMI index (p = 0.032); and negatively with iceA1 (p = 0.049) and peptic ulcer with cagA (p = 0.018). Thirty-five H. pylori strains were cultured and tested by E-test method showing that 49% of strains were resistant to at least one of the tested antibiotics. This is the first study that reports the high incidence and diversity of allelic combination of virulence genes in gastroduodenitis patients in Poland. Genotyping of H. pylori strains confirmed the involvement of cagA gene and vacAs1m1 genotype in development and severity of gastric disorder.


2011 ◽  
Vol 2011 ◽  
pp. 1-5 ◽  
Author(s):  
Fariborz Mansour-Ghanaei ◽  
Omid Sanaei ◽  
Farahnaz Joukar

Background. We encountered repeatedly, in our clinical practice, discordant results between UBT and histopathology aboutH. pyloriinfection.Goal. To study the diagnostic accuracy of Heliprobe14C-urea breath test (14C-UBT) for detection ofH. pyloriinfection in an Iranian population.Study. We enrolled 125 dyspeptic patients in our study. All of them underwent gastroscopy, and four gastric biopsies (three from the antrum and one from the corpus) were obtained. One of the antral biopsies was utilized for a rapid urease test (RUT), and three others were evaluated under microscopic examination. Sera from all patients were investigated for the presence ofH. pyloriIgG antibodies. The14C-UBT was performed on all subjects using Heliprobe kit, and results were analyzed against the following gold standard (GS):H. pyloriinfection considered positive when any two of three diagnostic methods (histopathology, RUT, serology) are positive.Results. According to data analysis, the Heliprobe14C-UBT had 94% sensitivity, 100% specificity, 93% negative predictive value (NPV), 100% positive predictive value (PPV), and 97% accuracy, compared with GS.Conclusion. The Heliprobe14C-UBT is an easy-to-perform, rapid-response, and accurate test forH. pyloridiagnosis, suitable for office use.


2017 ◽  
Vol 20 (3) ◽  
pp. 491-499 ◽  
Author(s):  
M. Jankowski ◽  
J. Spużak ◽  
K. Kubiak ◽  
K. Glińska-Suchocka ◽  
M. Biernat

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.


2019 ◽  
Vol 13 (08) ◽  
pp. 720-726
Author(s):  
Diana F Rojas-Rengifo ◽  
Belen Mendoza ◽  
Carlos Jaramillo ◽  
Paula A Rodríguez-Urrego ◽  
José F Vera-Chamorro ◽  
...  

Introduction: The presence of H. pylori in the stomach is associated with gastric pathologies. However, its diagnosis through culture methods is challenging because of its complex nutritional requirements and microaerophilic conditions for optimal growth. The preferred method for rapid diagnosis of H. pylori is the Rapid Urease Test (RUT) from human biopsies, which relies on the high activity of the urease enzyme present in H. pylori. However, RUT cannot say much more information about H. pylori. This makes evident the need for bacterial culture to know essential information such as the strain type, the kind of infection present and the bacteria’s antibiotic susceptibility. Methodology: Gastric biopsies from 347 patients were used for H. pylori isolation. We correlated the culture results with the RUT and histological grading used at Hospital Universitario Fundación SantaFe de Bogotá (HU-FSFB), Colombia. The concordance between techniques was determined by the Cohen’s Kappa coefficient (K). The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were also calculated. Results: The culture standardization was successful, and it could be applied for diagnosis in the clinical practice. H. pylori was positive by culture in 88 (26.34%) patients. The concordance of RUT and culture was strong (K= 0.805), and between histology and culture was moderate (K= 0.763) as well as for the gold standard defined and culture (K= 0.80). Conclusions: We present evidence that RUT and histological methods will be better interpreted for diagnosis of H. pylori if combined with bacterial isolation in cholesterol enriched culture.


2018 ◽  
Vol 2018 ◽  
pp. 1-7 ◽  
Author(s):  
Ashwak M. F. Abu-Taleb ◽  
Randa S. Abdelattef ◽  
Amina A. Abdel-Hady ◽  
Farida H. Omran ◽  
Lobna A. El-korashi ◽  
...  

H. pylori infection causes peptic ulcer, chronic gastritis, mucosa-associated lymphoid tissue lymphoma, and gastric carcinoma. It has several virulence factors such as cytotoxin-associated gene A(cagA) and the induced by contact with epithelium antigen (iceA). We aimed to explore the relationship between cagA and iceA of H. pylori and gastrointestinal diseases. One hundred and eighteen patients who attended Gastrointestinal Endoscopy Unit at Zagazig University Hospitals, Egypt, were included in this study. Two gastric biopsies were collected and evaluated by rapid urease test (RUT) and PCR. cagA and iceA genes were amplified by PCR. We found that 54 patients (45.76%) were positive by both RUT and PCR. cagA and iceA genes were present in 57.4% and 46.29% of the studied patients, respectively. cagA was the most prevalent gene in gastritis (33.3%) and peptic ulcer (68.7%). iceA1/iceA2 positive genes were the most prevalent in gastric cancer (75%). iceA1 gene was present in 38.7% of cagA positive cases, but iceA2 gene was present in 45.2% of cagA positive cases. iceA1/iceA2 positive genes were present in 29% of cagA positive cases. In conclusion, cagA and iceA genes could be used as markers for severe gastrointestinal diseases. iceA gene was strongly related to cagA gene.


2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Antoine Abou Rached ◽  
Jowana Saba ◽  
Cesar Yaghi ◽  
Joyce Sanyour ◽  
Ahmad El Hajjar ◽  
...  

Helicobacter pylori (H. pylori) can cause a wide variety of illnesses such as peptic ulcer disease, gastric adenocarcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma. The diagnosis and eradication of H. pylori are crucial. The diagnosis of H. pylori is usually based on the rapid urease test (RUT) and gastric antral biopsy for histology. The aim of this study is to evaluate the numbers of needed biopsies and their location (antrum/fundus) to obtain optimal result for the diagnosis of H. pylori. Three hundred fifty consecutive patients were recruited, 210 fulfill the inclusion criteria and had nine gastric biopsies for the detection of H. pylori infection: two antral for the first RUT (RUT1), one antral and one fundic for the second (RUT2), one antral for the third (RUT3) and two antral with two fundic for histology (HES, Giemsa, PAS). The reading of the 3 types of RUT was performed at 1 hour, 3 hours and 24 hours and biopsies were read by two experienced pathologists not informed about the result of RUT. Results of RUT were considered positive if H. pylori was found on histology of at least one biopsy. The RUT1 at 1h, 3h and 24h has a sensitivity of 72%, 82% and 89% and a specificity of 100%, 99% and 87% respectively. The positive predictive value (PPV) was 100%, 99% and 85% respectively and the negative predictive value (NPV) of 81%, 87% and 90%. The RUT2 at 1h, 3h and 24h, respectively, had a sensitivity of 86%, 87% and 91% and a specificity of 99%, 97% and 90%. The PPV was 99%, 96% and 88% and NPV of 89%, 90%, 94%. The RUT3 at 1h, 3h and 24h, respectively, had a sensitivity of 70%, 74% and 84% and a specificity of 99%, 99% and 94%. The PPV was 99%, 99% and 92% and NPV of 79%, 81% and 87%. The best sensitivity and specificity were obtained for RUT1 read at 3h, for RUT2 read 1h and 3h, and the RUT3 read at 24h.This study demonstrates that the best sensitivity and specificity of rapid test for urease is obtained when fundic plus antral biopsy specimens are used with a reading time at 3 hours.


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