title: Enzyme linked aptamer assay: based on a competition format for sensitive detection of antibodies to Mycoplasma bovis in serum.
abstract: Mycoplasma bovis (M. bovis) is a major, but often overlooked, pathogen that causes respiratory disease, mastitis, and arthritis in cattle. It has been widespread in China since 2008. In this study, single-stranded DNA (ssDNA) aptamers with high affinity and specificity against the P48 protein of M. bovis were selected using microplates as the matrix. Of nine candidates, aptamer WKB-14 showed the best affinity in an indirect enzyme-linked aptamer assay (ELAA) and good specificity by dot blotting. To the best of our knowledge, this is the first time that an aptamer has been used in a competitive ELAA for the serological detection of M. bovis. The percent inhibition (PI) cutoff value of the indirect competitive ELAA (ic-ELAA) was 40%, assessed using 20 negative sera. In a comparative study of different detection methods, ic-ELAA with dc-ELISA and dot blotting had a higher positive detection rate than the other two commercial indirect ELISA kits.
authors.affiliation: Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University , Beijing, P. R. China.
authors.forename: Ping
authors.initials: P
authors.lastname: Fu
authors.forename: Zhenhong
authors.initials: Z
authors.lastname: Sun
authors.forename: Ziqiang
authors.initials: Z
authors.lastname: Yu
authors.forename: Yuewei
authors.initials: Y
authors.lastname: Zhang
authors.forename: Junjun
authors.initials: J
authors.lastname: Shen
authors.forename: Haiyan
authors.initials: H
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Xu
authors.forename: Fei
authors.initials: F
authors.lastname: Jiang
authors.forename: Huiling
authors.initials: H
authors.lastname: Chen
authors.forename: Wenxue
authors.initials: W
authors.lastname: Wu
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D052157:Aptamers, Nucleotide / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001667:Binding, Competitive; D002417:Cattle; D004277:DNA, Single-Stranded / Q000378:metabolism; D007124:Immunoenzyme Techniques / Q000379:methods*; D057230:Limit of Detection; D045682:Mycoplasma bovis / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded