title: Development of enzyme immunoassay for detection of DDT.
abstract: Dichlorodiphenyltrichloroethane (DDT) is one of the persistent organic pollutants (POPs) widely found in the environment and in the general population. In this study, a direct competitive enzyme immunoassay (EIA) has been developed for the quantitative analysis of DDT. To generate a specific polyclonal antibody for EIA, p, p'-DDT was conjugated to porcine thyroglobulin for rabbit immunization. At optimized EIA conditions, the standard curves ranged from 0.137 to 100 ng/mL with the quantification limit of 0.41 ng/mL. The coefficients of variation (CV%) were 5.42-10.53% for intra-assay and 6.04-7.26% for inter-assay. Cross-reactivities with DDT metabolites (DDTs, including o, p'-DDT, p, p'-DDD, o, p'-DDD, p, p'-DDE, o, p'-DDE, p, p'-dichlorobenzophenone (DCBP), o, p'-DCBP) were investigated. The polyclonal antibody showed relatively low and/or no cross-reactivity with these compounds, and the assay was seen to be highly selective for p, p'-DDT. Moreover, the DDTs could be ranked by their reactivity: DDT > DDD > DDE > DCBP. In addition, the characterization of the polyclonal antibody indicated that the antiserum possesses a high specificity for p, p'-isomers. The results indicated that the developed EIA using this antibody could be a convenient and supplemental analytical tool for monitoring DDT.
authors.affiliation: Faculty of Environmental and Symbiotic Sciences, Prefectural University of Kumamoto, 3-1-100 Tsukide, Kumamoto, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Hirano
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Kitamura
authors.forename: Ikuo
authors.initials: I
authors.lastname: Kato
authors.forename: Chizuko
authors.initials: C
authors.lastname: Yanaihara
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Iwamoto
authors.forename: Makiko
authors.initials: M
authors.lastname: Sekiyama
authors.forename: Chiho
authors.initials: C
authors.lastname: Watanabe
authors.forename: Takashi
authors.initials: T
authors.lastname: Nakamoto
authors.forename: Nobukazu
authors.initials: N
authors.lastname: Miyamoto
authors.forename: Yuta
authors.initials: Y
authors.lastname: Onishi
authors.forename: Koji
authors.initials: K
authors.lastname: Arizono
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D002347:Carps / Q000378:metabolism; D003429:Cross Reactions; D003634:DDT / Q000032:analysis*; D004784:Environmental Monitoring / Q000379:methods*; D004787:Environmental Pollution / Q000032:analysis*; D007124:Immunoenzyme Techniques / Q000379:methods / Q000592:standards; D007536:Isomerism; D010573:Pesticide Residues / Q000032:analysis*; D010575:Pesticides / Q000032:analysis*; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012989:Soil Pollutants / Q000032:analysis
chemical_list: D000906:Antibodies; D010573:Pesticide Residues; D010575:Pesticides; D012989:Soil Pollutants; D003634:DDT