The application of aptamer Apt-236 targeting PvpA protein in the detection of antibodies against Mycoplasma gallisepticum

Abstract

Mycoplasma gallisepticum (M. gallisepticum) is the primary agent of chronic respiratory disease causing important economic losses in the poultry industry. Compared to antibodies, aptamers used to diagnose M. gallisepticum have many advantages, such as being chemically, animal-free produced and easily modifiable without affecting their affinity. Herein, a single-stranded DNA (ssDNA) aptamer Apt-236 which can specifically bind to PvpA protein of M. gallisepticum with a K_d of 1.30 ± 0.18 nM was selected successfully. An indirect blocking ELAA (ib-ELAA) for M. gallisepticum antibodies detection was also developed using Apt-236, in which M. gallisepticum antibodies would block the binding-position of aptamers. Therefor positive sera would prevent color development whereas negative sera will allow a strong color reaction. The ib-ELAA was consistent with other three widely used assays in terms of the growth and decline of the antibody response to M. gallisepticum, and showed substantial agreement with the results obtained using a commercial ELISA kit in clinical chicken sera samples. Therefore, the ib-ELAA developed in this study was a new format for aptamer application and would be an alternative method for the surveillance of M. gallisepticum.