Role of 13C-urea breath test in experimental model of Helicobacter pylori infection in mice

Background: Animal models have been widely used to study Helicobacter pylori infection. Evaluation of H. pylori infection status following experimental inoculation of mice usually requires euthanasia. The ¹³C‐urea breath test (¹³C‐UBT) is both sensitive and specific for detection of H. pylori in humans. Thus, it would be very useful to have such a test with the same accuracy for the follow‐up of this infection in animal models of gastric infection. Accordingly, the purpose of this study was to develop and evaluate a ¹³C‐UBT method for following the course of H. pylori infection in a mouse model. Material and Methods: A total of 50 female C57BL/6 mice were gavaged three times with either 10⁸ colony‐forming units of H. pylori (n = 29) or saline solution only (n = 21). After 2 months of infection, mice were fasted for 14 hours and ¹³C‐UBT was performed using 300 μg of ¹³C‐urea. The mice were killed, and the stomach was removed and processed for immunohistochemistry and PCR. Results: The optimal time for breath sample collection in mice was found to be 15 minutes. The ¹³C‐UBT cutoff was set at 3.0‰δPDB. Using PCR as the gold standard, the sensitivity of ¹³C‐UBT and immunohistochemistry was 96.6 and 72.4%, respectively, while the specificity was 85.7 and 95.2%, respectively. Conclusions: ¹³C‐UBT was shown to be a reliable method for the detection of H. pylori infection in C57BL/6 mice and was even more accurate than immunohistochemistry. The use of ¹³C‐UBT in the mouse model of H. pylori infection can be very useful to detect the bacterium without the need to kill the animals in long‐term time course studies.     

13C-urea breath test for the diagnosis of Helicobacter pylori infection in children: a systematic review and meta-analysis

Background: The ¹³C‐urea breath test (¹³C‐UBT) is a safe, noninvasive and reliable method for diagnosing H. pylori infection in adults. However, the test has shown variable accuracy in the pediatric population, especially in young children. We aimed to carry out a systematic review and meta‐analysis to evaluate the performance of the ¹³C‐UBT diagnostic test for H. pylori infection in children. Methods: We conducted a systematic review of the PubMed, Embase and Liliacs databases including studies from January 1998 to May 2009. Selection criteria included studies with at least 30 children and reporting the comparison of ¹³C‐UBT against a gold standard for H. pylori diagnosis. Thirty‐one articles and 135 studies were included for analysis. Children were stratified in subgroups of <6 and ≥6 years of age, and we considered variables such as type of meal, cutoff value, tracer dose, and delta time for the analysis. Discussion: The ¹³C‐UBT performance meta‐analyses showed 1, good accuracy in all ages combined (sensitivity 95.9%, specificity 95.7%, LR+ 17.4, LR? 0.06, diagnostic odds ratio (DOR) 424.9), 2, high accuracy in children >6 years (sensitivity 96.6%, specificity 97.7%, LR+ 42.6, LR? 0.04, DOR 1042.7), 3, greater variability in accuracy estimates and on average a few percentage points lower, particularly specificity, in children ≤6 years (sensitivity 95%, specificity 93.5%, LR+ 11.7, LR? 0.12, DOR 224.8). Therefore, the meta‐analysis shows that the ¹³C‐UBT test is less accurate for the diagnosis of H. pylori infection in young children, but adjusting cutoff value, pretest meal, and urea dose, this accuracy can be improved.     

体外~(14)C-尿素呼气试验检测胃内幽门螺杆菌感染研究

目的　建立诊断胃内幽门螺杆菌感染 (Hp)的体外 1 4 C-尿素呼气试验 (1 4 C- U BT)。方法　 47例 Hp阳性和 32例 Hp阴性患者接受测试 ,用口服微量胃液采集胶囊的办法收集胃液标本于一 10 m l无菌试管内 ,加入生理盐水 0 .5 m l和 18.5 k Bq1 4 C-尿素后立即加橡皮塞密封试管 ,室温放置反应 3h,注射器经橡皮塞注入 2 M H2 SO41.0 ml,使 1 4 CO2 释出。同一注射器回抽气体并立即注入装有 6 .5 ml的 1 4 CO2 搜集闪烁剂液闪瓶内搜集 1 4 CO2 ,最后在液体闪烁计数仪上作 1 4 C放射性测定。结果　 47例 Hp阳性病人 1 4 C放射性几何均数为 5 30 dpm,而 32例 Hp阴性者结果为 2 1dpm,二者相差 2 3倍 (Wilcoxon秩和检验 ,u=5 .5 976 ,P<0 .0 1)。以受试者工作特征曲线分析法得出判别阈值为 75 dpm ,对 Hp诊断的敏感性和特异性为 92 %(4 3/ 47)和 91% (2 9/ 32 )。结论　体外 1 4 C- UBT诊断 Hp感染具有高度的准确性 ,无放射性损伤之虞 ,可适用于临床诊断。     

First-time urea breath tests performed at home by 36,629 patients: a study of Helicobacter pylori prevalence in primary care

Background: The aim of the current study was (1) to describe the use of a ¹³C‐urea breath test (UBT) that was performed by patients at their homes as a part of a test‐and‐treat strategy in primary care and (2) to investigate the prevalence of Helicobacter pylori in patients taking a first‐time UBT. Material and Methods: The patients performed UBTs at home based on the discretion of the general practitioner and mailed the breath bags to a central laboratory for analysis. Each patient was identified by a unique civil registration number. The study was population‐based, and the background population was approximately 700,000 people. Results: From 2003 to 2009, 44,487 UBTs were performed. Of these, 36,629 were first‐time UBTs. In total, 726 of 45,213 breath bags received (1.6%) were unable to be analyzed because of errors with the bags. For both women and men who were ≤45 years of age, positive H. pylori declined over the time course of the study (women: 19.6% in 2003 to 17.6% in 2009, p < .01; men: 20.7% in 2003 to 16.9% in 2009, p < .001). Patients who were older than 45 years had significantly higher positive H. pylori results than younger patients. Conclusions: A test‐and‐treat system was possible to implement that allowed patients to perform UBTs at their homes. The results of the first‐time UBTs demonstrated that approximately one of five patients who presented with dyspepsia in the clinical setting of Danish primary care was infected with H. pylori.     

Problem of distinguishing false-positive tests from acute or transient Helicobacter pylori infections

BACKGROUND: Reliable detection of acute Helicobacter pylori infections remains problematic. The high prevalence of false-positive non-invasive tests in low H. pylori prevalence populations makes identification of acute and transient infections difficult. METHODS: We explored the use of serum pepsinogens (PG) for diagnosis of acute infection in patients following H. pylori challenge such that the onset of the infection was known. We then compared those findings to a group of children with presumed acute infections defined as a positive urea breath test (UBT) and negative IgG serology. RESULTS: We examined the pattern and calculated cut-off values of PG levels in 18 adult volunteers with known acute H. pylori infection. We then compared the results with sera from nine symptomatic children with presumed acute H. pylori infection and a matched control group of nine children who did not meet criteria for acute H. pylori infection. In acute infection, both PGI and II levels increased following H. pylori infection reaching a peak by 2 weeks post-infection. The frequency of a positive test defined as a value > mean +2 SD was 17, 71, and 94% at week 1, 2, and 4 post-infection, respectively. Only one child with presumed acute H. pylori infection had an elevated serum PGI and one had an elevated PGII. Five of the children had follow-up UBTs and four were negative consistent with the diagnosis of false-positive UBT. H. pylori infection was confirmed in the child with an elevated PGI level. CONCLUSIONS: These data suggest that a single positive noninvasive test in populations of low prevalence is most likely a false-positive result. This suggests that a single positive test requires confirmation preferably using a test that measures a different parameter (e.g., UBT confirmed by stool antigen test). It appears that most "transient"H. pylori infections are diagnosed on the basis of false-positive tests. PG levels are possible candidates as the confirmatory test.     

Does the Diagnostic Accuracy of the 13C‐Urea Breath Test Vary with Age Even After the Application of Urea Hydrolysis Rate?

Background: Endogenous CO₂ production may be a possible explanation for higher false‐positive results reported for ¹³C‐urea breath test (UBT) in children below 6 years. In this study, we evaluated whether age affects the diagnostic accuracy of the ¹³C‐UBT even after the application of urea hydrolysis rate (UHR) in children. Methods: A total of 612 ¹³C‐UBTs and endoscopic biopsies were performed on children divided into two groups; children under 6 years (n = 126) and children aged 6–18 years (n = 486). For ¹³C‐UBT, 75 mg ¹³C‐urea was ingested, and breath sample was collected 30 minutes later. Delta over baseline (DOB) was determined, and UHR was calculated to normalize the DOB values for endogenous CO₂ production. Results: There was significant difference between the DOB values of children under 6 years and those of children over 6 years in H. pylori‐positive (p = .029) and ‐negative groups (p = .002). On applying the UHR, no significant difference was observed between the UHR values of children under 6 years and those of children over 6 years in H. pylori‐positive (p = .877) and ‐negative groups (p = .427). In 12.6% children under 6 years, false‐positive results were observed on applying the DOB, and in 9.0% on applying the UHR (p = .125). Conclusions: The ¹³C‐UBT is a noninvasive method exhibiting high diagnostic accuracy with both UHR as well as DOB. However, high false‐positive results for ¹³C‐UBT were noted in children below 6 years on applying both UHR as well as DOB. Thus, this may not only be due to the effects of endogenous CO₂ production but also due to other factors.     

An evaluation of a serologic test with a current infection marker of Helicobacter pylori before and after eradication therapy in Chinese

Background and Aims: Development of an accurate and less cumbersome noninvasive method to detect current Helicobacter pylori infection is essential in clinic. The aim of this study was to evaluate the performance of the CIM test, also known as the Assure®H. pylori Rapid Test (Genelabs Diagnostics Pty. Ltd., Singapore), for the diagnosis of current H. pylori infection before and after eradication therapy in Chinese population. Methods: A total of 452 eligible people were recruited for this study in Jiangsu Province, China. Each individual underwent a ¹³C urea breath test (¹³C‐UBT). For the evaluation of CIM test after eradication, 115 H. pylori‐positive outpatients were treated with 1‐week triple therapy. One month after the end of therapy, the patients underwent ¹³C‐UBT again, and the CIM‐test was performed 1, 3, and 6 months after the end of therapy. Its performance (sensitivity, specificity, positive and negative predictive values, and accuracy) were determined using the ¹³C‐UBT as a gold standard for H. pylori diagnosis. Results: H. pylori was detected in 221 (65.6%) of the 337 people by ¹³C‐UBT. The sensitivity, specificity, positive and negative predictive values, and accuracy of the CIM test were 93.2%, 90.5%, 94.9%, 87.5%, and 92.3%, respectively, using ¹³C‐UBT as a gold standard. One month after eradication therapy, the sensitivity, specificity of CIM test were only 50% and 66.7%, 66.7% and 84.6% 3‐month after eradication therapy and the sensitivity, specificity increased to 85.7% and 96.9%, respectively, when CIM test was used 6 months after the end of anti‐H. pylori therapy. Conclusions: The CIM test is a simple, rapid, accurate, cheap, and near‐people test. It may be satisfactory for detecting H. pylori infection in cases without eradication therapy, but it could not differentiate the past or current infection correctly within 6 months after anti‐H. pylori therapy.