A Specific Method for the Detection of Hiochi Bacteria Using an Enzyme-linked Immunosorbent Assay System with Anti-hiochi Bacteria Monoclonal Antibodies.

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Ten standard strains of hiochi bacteria were selected based on the SDS-PAGE patterns of their cellular proteins. We then obtained ten hybridoma systems that secreted highly reactive monoclonal antibodies (MAbs) to the whole cells of each strain. It was apparent that these MAbs were highly reactive and specific for hiochi bacterial whole cells when using an enzyme-linked immunosorbent assay (ELISA). Three MAbs (two against the homo-fermentative hiochi lactobacilli and an MAb against the hetero-fermentative true hiochi bacilli) showed cross-reactivity to some of the other strains of lactobacilli tested. However, the other MAbs did not react with strains other than the immunogen. A sensitive ELISA method for the detection of hiochi bacteria was examined. It was possible to detect the order of 103 cells of the ten standard strains. Using this procedure and a mixture of the ten MAbs, a detection limit of 104 cells or less could be obtained for 98.2% of the hiochi bacteria isolated from sake brewing factories. Thus, this immunological technique using MAbs specific for hiochi bacteria is a sensitive and rapid detection method for hiochi bacteria, which can be used in the quality control of sake.

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