Development of an immunochromatographic lateral flow device for rapid detection of Helicobacter pylori stool antigen

Objective: Helicobacter pylori stool antigen (HpSA) test is a convenient and reliable non-invasive diagnosis for H. pylori infection. The aim of this study is to develop an immunochromatographic testing device for rapid detection of HpSA among Chinese patients. Design and methods: Monoclonal antibodies (McAb) targeting H. pylori were developed by conventional methods and paired by sandwich ELISA. The lateral flow device (LFD) was prepared using the selected McAb pair. A total of 867 clinically separated bacterial strains, including 56 H. pylori strains, were employed to test the sensitivity and specificity. Subsequently, the LFD was used to test 1200 human fecal samples, with a commercial HpSA testing device as comparison. Results: Two McAb pairs targeting H. pylori, DF2a/EE10b and IH10b/EE10b, were developed and proven to be of high specificity and sensitivity. After testing the cultures of 56 clinically separated H. pylori strains, the final LFD product was prepared using the mixture of DF2a and IH10b as capture antibodies and EE10b as the detection antibody. The testing threshold for H. pylori culture was 1.0 x 10(4) cfu/mL. The sensitivity and specificity were both 100% for the 867 tested bacterial cultures. The testing results of 1200 fecal samples showed that the positive and negative agreement rates between the homemade LFD and the commercial testing device were 99.75% and 99.87%, respectively. Conclusion: Our homemade HpSA LFD can be a very promising testing device for rapid diagnosis and epidemic screening of H. pylori infection among Chinese patients. (C) 2015 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.