A Unique and Sensitive ELISA Technique for Typing ABH Antigens in Bloodstains Using UEA-I Lectin—The Removal of Detergent with a Sephadex G-25 Mini-Column Improves Sensitivity

    Volume 41, Issue 1 (January 1996)

    ISSN: 0022-1198

    CODEN: JFSOAD

    Page Count: 5


    Nakamura, H
    Chief Scientist, Criminal Investigation Laboratory, Shimane Prefectural Police Headquarters, Matsue,

    Yuasa, I
    Assistant Professor, Tottori University School of Medicine, Yonago,

    Kimura, K
    Associate Professor, Research Assistant, Research Associates and Professor, Department of Legal Medicine, Central Research Laboratories, Shimane Medical University, Izumo,

    Matsubara, K
    Associate Professor, Research Assistant, Research Associates and Professor, Department of Legal Medicine, Central Research Laboratories, Shimane Medical University, Izumo,

    Tanabe, K
    Associate Professor, Research Assistant, Research Associates and Professor, Department of Legal Medicine, Central Research Laboratories, Shimane Medical University, Izumo,

    Takahashi, S
    Associate Professor, Research Assistant, Research Associates and Professor, Department of Legal Medicine, Central Research Laboratories, Shimane Medical University, Izumo,

    Idzu, T
    Associate Professor, Research Assistant, Research Associates and Professor, Department of Legal Medicine, Central Research Laboratories, Shimane Medical University, Izumo,

    Tanabe, Y
    Instructor, Central Research Laboratories, Shimane Medical University, Izumo,

    (Received 10 February 1995; accepted 19 June 1995)

    Abstract

    A unique sandwich enzyme-linked immunosorbent assay (ELISA) for the determination of ABH antigens in bloodstains has been developed. Human anti-A and -B antisera and Ulex europaeus anti-H lectin were coated on the inner surface of microplate wells. The sample antigens from bloodstains, solubilized with n-octyl-β-D-glucopyranoside which was then removed by passing through a Sephadex G-25 (G-25) mini-column, were placed in the wells. After washing the wells repeatedly, peroxidase-conjugated Ulex europaeus lectin I was added and incubated. Antigen activities were determined by the development of colors using o-phenylenediamine/H2O2. This technique permitted clear detection of all ABH antigens corresponding to the antisera and lectin with high sensitivities. The A and B antigens were solubilized as aggregates with H antigen from the erythrocyte membrane. Excess detergent remaining in the sample reduced the sensitivity and accuracy of this ELISA, probably due to the removal of antibody from the wells by the effect of the surfactant. The treatment of solubilized antigens with G-25, an indispensable step, eliminated the adverse effect of the detergent on the ELISA. The ELISA method reported here was proved to be easy, economical and sensitive, and this technique should be useful in the forensic practice.


    Paper ID: JFS13893J

    DOI: 10.1520/JFS13893J

    ASTM International
    is a member of CrossRef.

    Author
    Title A Unique and Sensitive ELISA Technique for Typing ABH Antigens in Bloodstains Using UEA-I Lectin—The Removal of Detergent with a Sephadex G-25 Mini-Column Improves Sensitivity
    Symposium , 0000-00-00
    Committee E30