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KMID : 0921620180480010014
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Journal of Bacteriology and Virology
2018 Volume.48 No. 1 p.14 ~ p.22
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Characterization of Specific IgA Response to Antigenic Determinants of Helicobacter pylori Urease Encoded by ureA and ureB in Children
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Shin Min-Kyoung
Jun Jin-Su
Kwon Soon-Wook
Lee Dong-Hae
Ha Jong-Hun
Park Jin-Sik
Song Dae-Hyun
Jung Myung-Hwan
Kang Hyung-Lyun
Park Ji-Sook
Youn Hee-Shang
Cho Myung-Je
Seo Ji-Hyun
Lee Woo-Kon
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Abstract
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Helicobacter pylori (H. pylori ), a causative agent of chronic gastritis and gastric cancer, has several virulent factors for own survival and progression toward gastric diseases in human stomach. Of those, H. pylori produces mainly urease (10~15% total protein weight) that neutralize the gastric acid for survival. Here, we identified the antigenic epitope of urease and then developed an ELISA using the antigen including the epitope of urease. We identified the antigenic epitope of urease that induces IgA antibodies in human using truncated mutants. Eight kinds of serially-truncated mutant of UreA and UreB were prepared and subjected to immunoblot using pooled sera of patients with gastric disorders. UreBEnd protein containing UreB epitope was produced and investigated its diagnostic value via ELISA in children. As a result, mutants having last 24 amino acid residues of UreB carboxyl terminus deleted did not show IgA-reactive band. The clones that contained the downstream of 448th amino acid in UreB showed IgA-reactive band. The serodiagnostic value of the UreBEnd recombinant protein including identified epitope was confirmed via IgA ELISA and shown to have 97% sensitivity and 100% specificity. These results demonstrated that carboxyl terminal region of UreB carries an antigenic epitope for IgA response in human. It may be useful for detecting H. pylori infection with improved test accuracy and minimum use of endoscopy.
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KEYWORD
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Helicobacter pylor, Urease, Epitope, IgA, ELISA, Children
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