Enzyme Linked Aptamer Assay: Based on a Competition Format for Sensitive Detection of Antibodies to Mycoplasma bovis in Serum

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• 作者：Ping Fu ; Zhenhong Sun ; Ziqiang Yu ; Yuewei Zhang ; Junjun Shen ; Haiyan Zhang ; Wei Xu ; Fei Jiang ; Huiling Chen ; Wenxue Wu
• 刊名：Analytical Chemistry
• 出版年：2014
• 出版时间：February 4, 2014
• 年：2014
• 卷：86
• 期：3
• 页码：1701-1709
• 全文大小：369K
• ISSN：1520-6882

文摘

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.