• Title/Summary/Keyword: H. pylori culture

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Expression of Helicobacter pylori Urease Subunit A in Plant (Helicobacter pylori urease subunit A 단백질의 식물체내에서의 발현)

  • 이효정;이만형;신동일;정일경;최성진;박희성
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.239-243
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    • 2000
  • Helicobacter pylori, an etiologic agent of gastritis and peptic ulceration, produces urease which elicits a powerful immunoglobulin response in H. pylori-infected individuals. To establish a model plant vaccine agains H. pylori, 750 bp -ureA DNA amplified by polymerase chain reaction from pH 808 plasmid harboring urease gene cluster was cloned and manipulated to be expressed in tobacco plants. From the regenerated transgenic tobacco plants, ureA DNA integration,m its mRNA expression and protein synthesis were analyzed and confirmed by standard molecular techniques. The CaMV 35S promoter-driving ureA construct was expressed to produce a 30 kDa protein which was identical with bacterial UreA in size when detected on immunoblot of SDS polyacrylamide gel electrophoresis.

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Relationship between Eradication of Helicobacter pylori Infection and Clarithromycin Resistance (Helicobacter pylori 감염의 치료와 Clarithromycin 내성간의 연관성)

  • Sohn Seung Ghyu;Lee Jong Hwa;Lee Jung Hun;Lee Sang Hee
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.177-182
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    • 2005
  • H. pylori strains were isolated from antral biopsies taken during upper endoscopy in 114 dyspeptic patients with no previous therapy against H. pylori. Rapid urease test, PCR amplification of SSA and cagA gene for H. pylori detection, and Western blot for CagA expression detection were performed. H. pylori infected patients were treated with omeprazole, clarithromycin (a macrolide), and amoxicillin. At 6 weeks after the discontinuation of therapy, the bacterial eradication rate was determined by endoscopy. The resistance rate to clarithromycin and amoxicillin was $20.2\%$ and $0.0\%$, respectively. The clarithromycin resistance was mainly caused by the A2142G mutation in the 23S rRNA gene of H. pylori. MICs of clarithromycin for the A2142G mutant isolates were significantly higher than MICs for the A2143G mutant isolates. H. pylori eradication was obtained in all patients with clarithromycin-susceptible isolates but not in patients with clarithromycin-resistant isolates (P = 0.0001). These results did not appear to be biased by any differences in CagA expression. The resistance of H. pylori to clarithromycin included in the therapeutic regimens is the most important reason for treatment failure. H. pylori antimicrobial susceptibility testing of the gastric biopsy culture should be performed before choosing the first triple therapy in infected patients and the increase in prevalence of clarithromycin resistance in Korea was problematic.

Evaluation of Invasive and Noninvasive Methods for the Diagnosis of Helicobacter Pylori Infection

  • Cosgun, Yasemin;Yildirim, Abdullah;Yucel, Mihriban;Karakoc, Ayse Esra;Koca, Gokhan;Gonultas, Alpaslan;Gursoy, Gul;Ustun, Huseyin;Korkmaz, Meliha
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.12
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    • pp.5265-5272
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    • 2016
  • Objective: The present study was conducted to evaluate invasive and noninvasive diagnostic methods for detection of Helicobacter pylori (H. pylori) in patients admitted with dyspeptic complaints and to compare sensitivities and specificities. Method: Sets of four gastric biopsy specimens were obtained from a total of 126 patients included in the study. The presence of H. pylori was determined by invasive tests including culture, rapid urease test, polymerase chain reaction (PCR) and histopathology. Among noninvasive tests, urea breath test, serological tests and enzyme-linked immunosorbent assay (ELISA) were performed. Results: H. pylori was isolated in 79 (62.7%) gastric biopsy cultures, whereas positivity was concluded for 105 (83.3%) patients by rapid urease test, for 106 (84.1%) by PCR, for 110 (87.3%) by histopathology, for 119 (94.4%) by urea breath test, and for 107 (84.9%) by ELISA. In the present study, the culture findings and histopathological examination findings were accepted as gold standard. According to the gold standard, urea breath test had the highest sensitivity (96.5%) and the lowest specificity (30%), whereas culture and histopathology had the highest specificities (100%). Conclusion: The use of PCR invasively with gastric biopsy samples yielded parallel results with the gold standard. PCR can be recommended for routine use in the diagnosis of H. pylori.

Helicobacter pylori Infection Impacts on Functional Dyspepsia in Thailand

  • Piriyapong, Kitiyawadee;Tangaroonsanti, Anupong;Mahachai, Varocha;Vilaichone, Ratha-Korn
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10887-10891
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    • 2015
  • Background: Helicobacter pylori (H. pylori) is a well known major cause of gastric cancer and even when asymptomatic infected patients are at elevated risk. Functional dyspepsia (FD) is also one of the most common gastrointestinal diseases, which greatly impacts the quality of life. H. pylori infection and psychosocial stress are frequently associated with FD but limited studies have confirmed the relationships, especially in Southeast Asian countries. Here we aimed to investigate the prevalence and impact of H. pylori infection, anxiety and depression on Thai FD patients. Materials and Methods: This cross-sectional study was conducted in a tertiary care center in Thailand, during February 2013-January 2014. All FD patients were diagnosed and categorized by Rome III criteria into epigastric pain syndrome (EPS) and postprandial distress syndrome (PDS) groups. The Hospital Anxiety and Depression Scale was used to evaluate psychological status. The presence of H. pylori was defined as positive with H. pylori culture, positive rapid urease test or positive histology. Results: Three hundred FD patients were included, 174 (58%) female. Overall mean age was 54.8+15.1 years. There were 192 (64%) patients with PDS and 108 (36%) with EPS. H. pylori infection was demonstrated in 70 (23.3%) patients. Anxiety and depression were documented in 69 (23%) and 22 (7.3%), respectively. H. pylori infection, anxiety and depression were significantly higher in PDS than EPS patients (27.1% vs 16.7%; p=0.04; OR=1.86; 95%CI=1.01-3.53 and 29.7% vs 11.1%; p=0.0002; OR=3.4; 95%CI=1.7-7.1 and 10.4% vs 1.9%; p=0.006; OR=6.2; 95%CI=1.4-38.9, respectively). Conclusions: H. pylori infection, anxiety and depression were commonly found in Thai FD patients and more prevalent in PDS than EPS. H. pylori eradication might be the key to success for the treatment of Thai FD patients and prevent the development of gastric cancer.

Antimicrobial Effect of Lactic Acid Bacteria Isolated from Kimchi and Tarak on Helicobacter pylori (김치와 타락에서 분리한 젖산균의 Helicobacterpylori에 대한 항균 효과)

  • Lee, Young-Duck;Yoo, Hye-Lim;Hwang, Ji-Yeon;Han, Bok-Kyung;Choi, Hyuk-Joon;Park, Jong-Hyun
    • The Korean Journal of Food And Nutrition
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    • v.23 no.4
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    • pp.664-669
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    • 2010
  • Lactic acid bacteria from traditional Korean foods of Tarak and Kimchi was isolated and characterized against carcinogenic Helicobacter pylori. Five Tarak and 30 Kimchi, traditional lactic acid-fermented foods, were collected from Andong area and the markets in Seoul, respectively and 15 lactic acid bacteria were isolated. Among them, two isolates were selected from high growth-inhibitory activities on H. pylori. The isolates were identified as Streptococcus thermophilus LAB kw15 from Tarak and Leuconostoc mesenteroides LAB kw5 from Kimchi by the biochemical characteristics and 16S DNA sequencing. The culture solutions of the isolates adjusted to pH 7.0 showed H. pylori inhibition. The isolates grew well and H. pylori did not grow during the co-culture with those strains. Therefore, L. mesenteroides LAB kw5 and S. thermophilus LAB kw15 might be the candidates as the functional lactic acid bacteria for improving stomach health.

Antagonism against Helicobacter Pylori and Proteolysis of Lactobacillus Helveticus CU631 and Strain Identification

  • Yoon, Y.H.;Won, B.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1057-1065
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    • 2002
  • The antagonistic activities of 30 strains of lactobacilli against Helicobacter pylori were determined and Lactobacillus helveticus CU631 has been selected as the strain which possesses the strongest inhibitory effect in the disc diffusion assay showing inhibition zone diameter of $10{\pm}1.5mm$, whereas those of L. plantarum and L. fermentum have been shown to be $4.0{\pm}0.6mm$. H. pylori G88016 revealed the highest vacuolating toxin producing activity among the 8 strains, the inhibitory activity of L. helveticus CU631 in vacuolating toxin producing activity of H. pylori manifested in the co-culture of two strains and in the 5:5 mixture of supernatant of the two strains. Both L. helveticus CU631 and cell free culture supernatant had a strong inhibitory activities in urease and cytotoxin producing activities of H. pylori NCTC11637 and CJH12. An accelerated proteolytic activity of water soluble peptides by L. helveticus CU631 during the refrigeration storage has been manifested in the cream cheese. DNA seqences of 16S-23S ribosomal RNA spacer region showed typical pattern among the various strains of L. helveticus, which could be used in the identification of L. helveticus CU 631.

Effect of Evodiae fructus Methanol Extract on Virulence-Related Genes' Expression of Helicobacter pylori (오수유 메탄올 추출물이 Helicobacter pylori 병원성 관련 유전자 발현에 미치는 영향)

  • Yang, Ji Yeong;Lee, Pyeongjae;Kim, Jong-Bae
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.316-322
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    • 2019
  • Helicobacter pylori, a gram-negative bacterium, is one of the risk factors that induces gastritis and gastric cancer. Therefore, much attention has been paid to the compounds that inhibit bacterial growth or eradicate bacteria. Evodiae fructus (EF), the fruit of Evodia rutaecarpa, has been used for treating diarrhea and abdominal pain. EF extract was already found to inhibit the growth of H. pylori. However, to the best of our knowledge, the effect of EF on the virulence factors of H. pylori has not been reported. In this study, when comparing the minimum inhibitory concentration (MIC) of the different methanol concentration extracts, the 95% methanol extract (EF95) showed the lowest MIC value. EF95 extract suppressed the expressions of cagA, vacA and ureB, but interestingly, it up-regulated the expression of ureA. A decrease in production of ammonia in the culture medium and the cell lysates indicated that EF95 inhibited the urease activity in H. pylori, which was the result of EF95 inhibiting the ureB expression. Although the mechanism by which EF95 extract regulates the virulence factors in H. pylori needs further study, EF95 could be used for treatment of gastric troubles induced by H. pylori.

Isolation and Characterization of Kimchi Lactic Acid Bacteria Showing Anti-Helicobacter pylori Activity (Helicobacter pylori 억제능 김치 유산균의 분리와 특성 규명)

  • Lee, Youl;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.36 no.2
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    • pp.106-114
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    • 2008
  • One bacterium, which showed strong antagonistic activity against H. pylori KCCM 41756, was isolated from kimchi. The strain NO1 was designated as Lactobacillus plantarum NO1 based on Gram staining, biochemical properties, and 16S rRNA gene sequencing. The culture medium $(2{\sim}4{\mu}g/ml)$ of Lb. plantarum NO1 reduced $(40{\sim}60%)$ the urease activity of H. pylori KCCM 41756. Lb. plantarum NO1 inhibited the binding of H. pylori to human gastric cancer cell line, AGS cells, by more than 33%. Lb. plantarum NO1 exhibited high viability (maintained initial viable cell count of $10^9CFU/ml$) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastricjuice for 2 h and in 0.3%, 0.5% oxgall for 24 h. Hemolysis phenomena did not observed when Lb. plantarum NO1 was incubated in the blood agar media. We concluded that Lb. plantarum NO1 can be a good candidate as a probiotic, harboring anti-H. pylori activity.

Anti-Helicobacter pylori Activity of Lactobacillus spp. Isolated from Gajami Sikhae (가자미식해에서 분리한 Lactobacillus spp.의 항헬리코박터 활성 평가)

  • Eun-Yeong Bae;Gi-Un Cho;Sung-Keun Jung;Young-Je Cho;Byung-Oh Kim
    • Journal of Life Science
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    • v.33 no.3
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    • pp.260-267
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    • 2023
  • Helicobacter pylori infects the mucosa, induces chronic inflammation and ulcers, and is known as a biological carcinogen. Antibiotics are used as therapeutic agents for H. pylori, but there are problems such as resistance. Thus, research is being conducted on the use of lactic acid bacteria (LAB) as an alternative therapeutic agent. There have been many studies on LAB related to kimchi. However, studies related to Gajami Sikhae, a traditional fermented seafood in Korea, are insufficient. In this study, we investigated the inhibitory effect of LAB isolated from Gajami Sikhae on H. pylori and its use as a probiotic. Forty species of LAB isolated from Gajami Sikhae were identified as Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc mesenteroides, and Weisella paramesenteroides, and 10 strains of 40 species were selected through liquid inhibition assay of H. pylori. The selected LAB supernatant at 1%, 5%, and 10% had a growth inhibitory effect on H. pylori 52, 51, e-53, and 309. The adjusted pH of 7.0 was used for the LAB culture supernatant, in reference to a study that the growth of H. pylori is affected by acid. All 10 strains of LAB at 5% and 10% concentration suppressed the growth of H. pylori 52, and 7 strains of LAB at 10% concentration suppressed the growth of H. pylori e-53. LAB also had the effect of suppressing the activity of urease. Finally, LAB isolated from Gajami Sikhae is expected to be useful for eradicating and preventing H. pylori.

Role of enzyme immunoassay for the Detection of Helicobacter pylori Stool Antigen in Confirming Eradication After Quadruple Therapy in Children (소아에서 4제요법 후 enzyme immunoassay에 의한 Helicobacter pylori 대변 항원 검출법의 유용성에 대한 연구)

  • Yang, Hye Ran;Seo, Jeong Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.7 no.2
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    • pp.153-162
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    • 2004
  • Purpose: The Helicobacter pylori stool antigen (HpSA) enzyme immunoassay is a non-invasive test for the diagnosis and monitoring of H. pylori infection. But, there are few validation studies on the HpSA test after eradication in children. The aim of this study was to assess the diagnostic accuracy of HpSA enzyme immunoassay for the detection of H. pylori to confirm eradication in children. Methods: From January 2001 to October 2003, 164 tests were performed in 146 children aged 1 to 17.5 years (mean $9.3{\pm}4.3$ years). H. pylori infection was confirmed by endoscopy-based tests (rapid urease test, histology, and culture). All H. pylori infected children were treated with quadruple regimens (Omeprazole, amoxicillin, metronidazole and bismuth subcitrate for 7 days). Stool specimens were collected from all patients for the HpSA enzyme immunoassay (Primier platinum HpSA). The results of HpSA tests were interpreted as positive for $OD{\geq}0.160$, unresolved for $$0.140{\leq_-}OD$$<0.160, and negative for OD<0.140 at 450 nm on spectrophotometer. Results: 1) One hundred thirty-one HpSA tests were performed before treatment. The result of HpSA enzyme immunoassay showed three false positive cases and one false negative case. The sensitivity, specificity, positive predictive value, and negative predictive value of HpSA enzyme immunoassay before treatment were 96.4%, 97.1%, 90%, and 99%, respectively. 2) Thirty-three HpSA enzyme immunoassay were performed at least 4 weeks after eradication therapy. The results of HpSA enzyme immunoassay showed two false positive cases and one false negative case. The sensitivity, specificity, positive predictive value, and negative predictive value after treatment were 88.9%, 91.7%, 80%, and 95.7%, respectively. Conclusion: Diagnostic accuracy of the HpSA enzyme immunoassay after eradication therapy was as high as that of the HpSA test before eradication therapy. The HpSA enzyme immunoassay was found to be a useful non-invasive method to confirm H. pylori eradication in children.

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