Development of easy, sensitive, and quantitative detection methods for enteric pathogens of word importance from food

2014 Fiscal Year Final Research Report

• Project/Area Number: 24249038
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hygiene
• Research Institution: Kyoto University
• Principal Investigator: NISHIBUCHI Mitsuaki 京都大学, 東南アジア研究所, 教授 (50189304)
• Co-Investigator(Kenkyū-buntansha): NAKAGUCHI Yoshitsugu 石川県立大学, 生物資源環境学部, 准教授 (70378967)
• Co-Investigator(Renkei-kenkyūsha): SETO Kazuko 大阪府立公衆衛生研究所, 主任研究員 (70211323)
• Project Period (FY): 2012-10-31 – 2015-03-31
• Keywords: 腸炎ビブリオ / 腸管出血性大腸菌 / 免疫磁気ビーズ法 / LAMP / IMS

Outline of Final Research Achievements

We have established a sensitive, easy, rapid, and quantitative method by incorporating an immunomagnetic method and a genetic method so-called the LAMP method to detect pathogenic (tdh- and/or trh-positive) strains of Vibrio parahaemolyticus. We demonstrated the dried-type LAMP reagent made possible the maintenance and transportation of the reagent and eliminated the need for gel electrophoresis to judge the result in the workshops held in seven developing countries in Asia and South America. We found loss of hemolytic activity of the trh gene product due to frequent base substitutions in this gene of environmental strains, and thus we are adjusting the LAMP primers for this gene to improve this detection method.
Regarding the enterohemorrhagic Escherichia coli, we developed a similar qualitative detection method for the strains belonging to 12 important O serotypes in the world from beef at once.

Free Research Field

病原細菌学