Production and characterization of a novel monoclonal antibody against Vibrio parahaemolyticus F₀F₁ ATP synthase's delta subunit and its application for rapid identification of the pathogen

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• 作者：Junko Sakata^a ; ^b ; ^{sakata@iph.pref.osaka.jp} ; Kentaro Kawatsu^a ; Tadashi Iwasaki^b ; Katsuhiro Tanaka^b ; Shigeo Takenaka^b ; Yuko Kumeda^a ; Hiroshi Kodama^b
• 关键词：Monoclonal antibody ; Dot blotting assay ; Vibrio parahaemolyticus ; F0F1 ATP synthase's delta subunit
• 刊名：Journal of Microbiological Methods
• 出版年：2012
• 出版时间：January 2012
• 年：2012
• 卷：88
• 期：1
• 页码：77-82
• 全文大小：401 K

文摘

We raised monoclonal antibodies (MAbs) against Vibrio parahaemolyticus cell extracts. One of the MAbs, designated MAb-VP34, reacted in enzyme-linked immunosorbent assays (ELISAs) with 140 V. parahaemolyticus strains, regardless of serotype or origin. MAb-VP34 did not detectably react with 96 strains belonging to 27 other Vibrio species (except for Vibrio natriegens) or with 29 non-Vibrio species. These results show that MAb-VP34 is highly specific for V. parahaemolyticus. Western blotting and mass spectrometry analyses revealed that MAb-VP34 recognized V. parahaemolyticus F₀F₁ ATP synthase's delta subunit.

Using MAb-VP34, a rapid and simple immunodot blotting assay (VP-Dot) was developed to determine whether bacterial colonies growing on selective agar, represented V. parahaemolyticus. To evaluate VP-Dot, 20 V. parahaemolyticus strains and 19 non-related strains were tested. The results indicated that VP-Dot is a reliable tool for identification of V. parahaemolyticus colonies. The simple VP-Dot procedure took 40 min, indicating that the MAb-VP34 based immunological method will greatly reduce labor, time, and costs required to verify V. parahaemolyticus colonies as compared with the conventional biochemical test.