Culture and successful eradication of Helicobacter pylori from heterotopic gastric mucosa.

Helicobacter pylori colonizes the gastric mucosa of humans and can cause chronic gastritis, peptic ulcer disease, gastric cancer or mucosa-associated-lymphoid tissue (MALT) lymphoma. Here, we report the case of a 61-year-old male patient who presented with tickle of the throat, globus sensation and heartburn. In an esophagogastroduodenoscopy subpharyngeal localized heterotopic gastric mucosa (HGM), reflux esophagitis and a chronic gastritis were diagnosed. HGM and stomach were H. pylori positive as proven by culture and histopathological examination. After eradication therapy with a proton pump inhibitor (PPI), amoxicillin and clarithromycin followed by PPI treatment, the patient reported clinical improvement and the histopathological changes in the HGM due to H. pylori infection improved, too. This case report demonstrates that culture and susceptibility testing of H. pylori using established protocols succeeds not only from tissue samples of the stomach but also from heterotopic gastric mucosa. Eradication therapy may not only improve typical H. pylori associated discomforts of the stomach but also extragastric signs and symptoms of H. pylori infection.

Is Helicobacter pylori resident or transient in the human oral cavity?

Helicobacter pylori colonizes the stomachs of at least half of the world's human population. The role of the oral cavity in this colonization is not clear and there are, to date, no comprehensive data that clearly demonstrate the isolation of this bacterium from the oral cavity. The aim of this study was to evaluate the prevalence of H. pylori in the oral cavity of 15 patients who tested positive for H. pylori. A comprehensive dental examination of all patients was conducted. Samples were taken from supragingival and subgingival plaque, saliva, periapical exudates and tongue swabs. All samples were taken before the application of antibiotics. A total of 163 oral samples were investigated by PCR using two different H. pylori-specific primer pairs. A PCR inhibition control using a modified plasmid was always included for the most specific primer pair. In addition, a culture technique was used to confirm PCR results. Despite a PCR detection limit of 10(2) bacteria ml(-1), out of 14 patients, H. pylori could not be detected in any of the samples taken. In one patient, H. pylori-positive PCR signals were obtained in two samples using only one primer pair. H. pylori could not be cultivated from these two PCR-positive samples; therefore, no correlation to oral colonization status could be established. This study challenges the misleading preconception that H. pylori resides in the human oral cavity and suggests that this bacterium should be considered transient and independent of the oral status. To date, positive PCR results for H. pylori in the oral cavity have been overestimated and not critically interpreted in literature.

[Networks of National Reference Centers and associated Consiliary Laboratories in Germany]. / Referenznetzwerke aus Nationalen Referenzzentren mit assoziierten Konsiliarlaboratorien in Deutschland.

Since 1995, the Robert Koch-Institute in agreement with the Federal Ministry of Health in Germany has increased the funding of National Reference Centers (NRC) and Consiliary Laboratories (CL) for laboratory-based surveillance of selected infection pathogens and infectious disease syndromes. Their goal is to improve efficient protection from infections and to supplement infectious disease surveillance by monitoring selected pathogens. Currently there are 19 NRC and 48 CL, nominated for a duration of 3 years. In order to enhance the effectiveness and cooperation of the system, ten National Networks were launched in 2009. The aim of these networks is to facilitate exchange on diagnostic methods and prevention concepts and to improve the geographic coverage of the services. Furthermore, the networks provide an opportunity to work on issues beyond single pathogens more productively and efficiently. In addition, the inclusion of external and international specialists should to be included more often in the future. The activities of the networks are evaluated by the commission for infectious disease epidemiology. The commission develops promotion modalities to support collaboration between NRC and CL and to adapt it to more closely meet the requirements at the national and international levels.

Seasonality of Clostridium difficile infections in Southern Germany.

Between 2000 and 2009, the total number of patients with Clostridium difficile infections increased considerably in Southeastern Germany. A clear seasonality was observed with a higher number of affected patients occurring in the winter months (January-March). Moxifloxacin and erythromycin-resistant C. difficile PCR ribotypes 001 (72%) and 027 (4·6%) were the most commonly isolated strains.

Recognition of Clostridium difficile PCR-ribotypes 001, 027 and 126/078 using an extended MALDI-TOF MS system.

During the last decade, Clostridium difficile infection (CDI) increased markedly inside as well as outside of hospitals. In association with the occurrence of new hypervirulent C. difficile strains, CDI became more important. Until now typing of C. difficile strains has been enabled by PCR-ribotyping. However, this method is restricted to specialized laboratories combined with high maintenance cost. Therefore, we tested MALDI-TOF mass spectrometry for typing of C. difficile to provide a fast method for surveillance of CDI. Using a standard set of 25 different C. difficile PCR ribotypes a database was made by different mass spectra recorded in the SARAMIS software (AnagnosTec, Zossen, Germany). The database was validated with 355 C. difficile strains belonging to 29 different PCR ribotypes collected prospectively from all submitted feces samples in 2009. The most frequent PCR ribotypes were type 001 (70%), 027 (4.8%) and 078/126 (4.7%). All three types were recognized by MALDI-TOF MS. We conclude that an extended MALDI-TOF system was capable to recognize specific markers for ribotypes 001, 027 and 078/126 allowing an effective identification of these strains.

Correlations between bed occupancy rates and Clostridium difficile infections: a time-series analysis.

A time-series analysis was performed to identify the impact of bed occupancy rates and length of hospital stay on the incidence of Clostridium difficile infections (CDI). Between January 2003 and July 2008, a mean incidence of 0·5 CDI cases/1000 patient days was recorded. Application of a multivariate model (R2=0·50) showed that bed occupancy rates on general wards (P<0·01) and length of stay in intensive care units (ICUs) (P<0·01) influenced the incidence of CDI. Overcrowding on general wards and long periods in ICUs were identified as being positively associated with the incidence of CDI.

[Emergence of clostridium difficile ribotype 027 in Germany: epidemiological and clinical characteristics]. / Clostridium-difficile-Ribotyp 027: Epidemiologie und Klinik des erstmaligen endemischen Auftretens in Deutschland.

INTRODUCTION: In September 2007 an increase of severe Clostridium difficile-associated infections (CDI) was noticed in a hospital in the city of Trier, Germany. It was assumed that a new, possibly hypervirulent strain (PCR ribotype 027) was related to these events. An outbreak investigation was initiated by the local health authorities and the Robert Koch Institute to describe the epidemiology of the possible outbreak and to identify and control the possible sources. METHODS: In addition to retrospective case-finding of severe CDI and ribotype 027 infections by analysis of patient documents and certificates of death, an active surveillance system for severe CDI and ribotype 027 infections was established in the 6 hospitals of the affected region. In all suspected cases, a test for toxin A/B and a stool culture for C. difficile were conducted simultaneously. Bacterial isolates were further characterised by PCR ribotyping. Data on the course of disease, case fatality, and possible risk factors for CDI-related deaths were assessed using a standardised questionnaire. Environmental investigations were done. RESULTS: By 31 January 2008, 27 cases of severe CDI and 21 cases with C. difficile ribotype 027 infections were found in the area under investigation. Active surveillance found 76 of 399 (19 %) patients positive for C. difficile. In 20 patients, PCR ribotyp 027 could be proven. In total, 9 deaths occurred (19 %). An existing immunosupressive therapy (OR 35.8; 95 % CI 2.8 - 464.5) was related to case fatality in the multivariate analysis. Severe cases of CDI were also observed in non-ribotype 027 infections. In the screening of hospital personnel (n = 161), 6 % were found positive for toxin A/B. DISCUSSION: This investigation demonstrated the endemicity of C. difficile PCR ribotype 027 in Germany for the first time. As a consequence from this study, severe CDI became a reportable disease in Germany at the end of 2007. In addition to hygienic measures, the critical use of antibiotics is an important measure to prevent a further increase of CDI.

Association of ciprofloxacin prescriptions to outpatients to Clostridium difficile infections.

To study if antibiotic treatment of outpatients had triggered Clostridium difficile infections (CDI), prescription numbers were compared with CDI-affected patient numbers. A strong correlation was observed for ciprofloxacin (R=0.917), suggesting that increased use of ciprofloxacin by outpatients contributed to increased numbers of CDI. These findings deserve further investigation as they may have an impact on future decisions regarding antibiotic prescribing.

S3-guideline "helicobacter pylori and gastroduodenal ulcer disease" of the German society for digestive and metabolic diseases (DGVS) in cooperation with the German society for hygiene and microbiology, society for pediatric gastroenterology and nutrition e. V., German society for rheumatology, AWMF-registration-no. 021 / 001.

This guideline updates a prior consensus recommendation of the German Society for Digestive and Metabolic Diseases (DGVS) from 1996. It was developed by an interdisciplinary cooperation with representatives of the German Society for Hygiene and Microbiology, the Society for Pediatric Gastroenterology and Nutrition (GPGE), and the German Society for Rheumatology. The guideline is methodologically based on recommendations of the Association of the Scientific Medical Societies in Germany (AWMF) for providing a systematic evidence-based S 3 level consensus guideline and has also implemented grading criteria according to the GRADE (Grading of Recommendations Assessment, Development, and Evaluation) process. Clinical applicability of study results as well as specifics for Germany in terms of epidemiology, antibiotic resistance status, diagnostics, and therapy were taken into account.

[Establishment of Networks of National Reference Centres and associated Consiliary Laboratories in Germany]. / Etablierung von Referenznetzwerken aus Nationalen Referenzzentren mit assoziierten Konsiliarlaboratorien in Deutschland.

The German Federal Ministry of Health has funded National Reference Centres (NRC) for laboratory-based surveillance of selected infection pathogens and infections disease syndromes. This selection is based on the epidemiologic relevance of the pathogens, specific diagnostic requirements, antimicrobial resistance and need for public health measures. Currently there are 18 NRC, nominated for a duration of 3 years. Toward the end of a nomination period, each NRC is evaluated by an expert committee, based on the catalogue of core tasks. In order to expand the spectrum of competencies 47 consiliary laboratories on additional pathogens of special epidemiologic importance have been named. Their main function is to provide information and consultation on special diagnostic issues. In order to further improve the effectiveness and cooperation of the system Networks have been created. The aim of the Networks is to facilitate exchange of diagnostic methods and prevention concepts and to improve the geographic coverage of the services.

[S3-guideline "Helicobacter pylori and gastroduodenal ulcer disease"]. / S3-Leitlinie "Helicobacter pylori und gastroduodenale Ulkuskrankheit" der Deutschen Gesellschaft für Verdauungs- und Stoffwechselkrankheiten (DGVS).

This guideline updates a prior concensus recommendation of the German Society for Digestive and Metabolic Diseases (DGVS) from 1996. It was developed by an interdisciplinary cooperation with representatives of the German Society for Microbiology, the Society for Pediatric Gastroenterology and Nutrition (GPGE) and the German Society for Rheumatology. The guideline is methodologically based on recommendations of the Association of the Scientific Medical Societies in Germany (AWMF) for providing a systematic evidence-based consensus guideline of S 3 level and has also implemented grading criteria according to GRADE (Grading of Recommendations Assessment, Development and Evaluation). Clinical applicability of study results as well as specifics for Germany in terms of epidemiology, antibiotic resistance status, diagnostics and therapy were taken into account.

Increased number of Clostridium difficile infections and prevalence of Clostridium difficile PCR ribotype 001 in southern Germany.

In recent years, Clostridium difficile infection (CDI) has emerged as an increasing problem, both in in- and outpatients. In a rural region of southern Germany, the annual number of C. difficile toxin (Tcd)-positive patients has increased from 95 to 796 in the period from 2000 to 2007. Simultaneously, the proportion of positive tests among all Tcd examinations has risen from 7.0% to 12.8%, indicating that the higher number of affected patients was not solely due to an increase in the number of assays. Elevated numbers of CDI have recently been associated with outbreaks of the ribotype 027 strain, particularly in North America. This strain has also been isolated in Europe, including in Germany. Ribotyping and PCR testing for binary toxin genes of C. difficile strains isolated from in- and outpatients demonstrate a predominance (59%) of C. difficile ribotype 001, which exhibits antibiotic resistance to erythromycin, ciprofloxacin, and moxifloxacin, but lacks binary toxin genes. In summary, in our region of Germany, the number of patients affected by CDI has increased, probably due to spread of C. difficile ribotype 001.

Antimicrobial resistance in Escherichia coli strains isolated from food, animals and humans in Germany.

In human medicine, increasing resistance in Escherichia coli strains causes therapeutic problems and is partly attributed to the selection pressure in veterinary medicine. The resistance proportions (RP) of E. coli isolates obtained in 2002/2003 from animals (n=110) and from food originating from animals (n=42) were highest for tetracyclines (57%), ampicillin (38%) and trimethoprim/sulfamethoxazole (37%). None of the strains tested positive for ESBL by VITEK. Comparison of Escherichia coli strains revealed higher RP in samples from humans for strains resistant to piperacillin, ampicillin/sulbactam and second generation quinolones, whereas E. coli strains originating from animals showed a higher resistance proportion to tetracycline.

[Pathogenesis, diagnostics and treatment of Helicobacter pylori infection]. / Pathogenese, Diagnostik und Therapie der Helicobacter-pylori-Infektion.

More than one-half of the world population is infected with Helicobacter pylori. Of those, approx. 500,000 die from gastric carcinoma every year. Ulcer disease, gastricatrophy and the rare MALT lymphoma are other sequelae of H. pylori infection. H. pylori possesses an array of virulence factors that include urease, flagellar motility, adhesins, the vacuolating cytotoxin VacA and the protein CagA. The gene encoding CagA is located on the cag pathogenicity island, comprising 29 genes the majority of which encodes components of a type IV secretion system capable of translocating CagA into epithelial cells where it interferes with cellular signal transduction processes. A number of diagnostic tests for H. pylori infection require gastroendoscopy. These include the biopsy urease test, histology, culture with susceptibility testing, and molecular detection methods such as fluorescent in situ hybridization. Non-invasive tests that do not require endoscopy include the (13)C urea breath test, H. pylori stool antigen ELISA and serology. The latter is unsuitable for treatment follow-up, since antibody titres persist up to a year after successful treatment. When patients have never been treated for H. pylori infection, biopsy urease test and histology are usually sufficient for diagnosis. In patients where endoscopy is not required, H. pylori infection can be reliably detected by (13)C urea breath test, stool antigen ELISA or serology. Patients who have under gone one or more unsuccessful cycles of eradication therapy in most cases harbour H. pylori resistant to one or several antibiotics. In these patients, culture and antibiotic susceptibility testing are indicated. Patients who have never been treated for H. pylori infection usually harbour susceptible strains. In such patients, classic "Italian" or "French" triple therapies may achieve eradication in >90% of cases. In the case of treatment failure, second-line antibiotic treatment regiments (rescue therapy) are used, optimally guided by susceptibility data.

Helicobacter pylori eradication and gastric ulcer healing--comparison of three pantoprazole-based triple therapies.

AIM: To study the efficacy of three pantoprazole-based triple therapy regimens for the eradication of Helicobacter pylori infection and gastric ulcer healing. METHODS: In an open, multi-centre, randomized study, 519 H. pylori-positive patients with active gastric ulcer were randomized to receive pantoprazole (40 mg) (P) and two of three antibiotics: clarithromycin (500 mg) (C), metronidazole (500 mg) (M) or amoxicillin (1000 mg) (A). Triple therapy (PAC, PCM, PAM) was administered twice daily for 7 days, followed by pantoprazole until the ulcer had healed. Antrum and corpus biopsies were taken to determine the pattern of gastritis, to assess the H. pylori status and to determine the strain susceptibility to antibiotics, and from the ulcer margins and base to exclude malignancy. Scores based on the Sydney system were used to categorize the gastritis phenotypically. RESULTS: The H. pylori eradication rates for the per protocol (intention-to-treat) analysis were 89% (67%) for PAC, 83% (68%) for PCM and 76% (60%) for PAM, with a significant difference between PAC and PAM. Healing rates after 4 weeks were 91% for PAM, 90% for PCM and 88% for PAC (per protocol analysis). The eradication rates were lower in patients in whom strains resistant to any antibiotic used in the triple therapies were detected. Successful eradication [odds ratio, 5.2 (3.3; 8.3)] and the ulcer size (< 15 mm) were significant predictors for healing after 4 weeks. The regimens showed a comparable safety profile and compliance. CONCLUSIONS: Pantoprazole-based triple therapies are effective in the eradication of H. pylori infection in gastric ulcer patients, as reported in previous similar sized studies in duodenal ulcer patients. Successful eradication and an ulcer size of < 15 mm are the best predictors of gastric ulcer healing after 4 weeks.

Antibiotic-associated diarrhea: therapeutic aspects and practical guidelines--an interdisciplinary approach to a common problem.

Antibiotic-associated diarrhea (AAD) is a common complication of antibiotic treatment, most often seen in non-hospitalised patients. In principle, such diarrhea can be triggered by any antibiotic. An interdisciplinary working group discussed the different aspects of AAD in view of its gastroenterological, microbiological, paediatric, general medical and pharmaceutical implications, also in consideration of the position of patients and health insurance funds. This paper implies therapeutic aspects and practical guidelines to raise awareness of these problems also in routine situations and to enable the persons and institutions involved on the various levels of the health-care system (patients, pharmacists, family doctors, specialists and hospitals) to handle the problem of AAD more easily in a standardised way as far as diagnostics, therapy and prevention are concerned.

Antibiotic-associated diarrhea: incidence, risk factors of antibiotics and patients, pathophysiology and differential diagnosis--an interdisciplinary approach to a common problem.

Antibiotic-associated diarrhea (AAD) is a common complication of antibiotic treatment, most often seen in non-hospitalised patients. In principle, such diarrhea can be triggered by any antibiotic. An interdisciplinary working group discussed the different aspects of AAD in view of its gastroenterological, microbiological, paediatric, general medical and pharmaceutical implications, also in consideration of the position of patients and health insurance funds. The incidence, risk factors of antibiotics and patients, the pathophysiology of the various types of AAD and the differential diagnosis are reviewed.