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Poor visualization of direct agglutination test (DAT) and low sensitivity of agar double immunodiffusion (ADID) test severely hindered their applications in large-scale sample screening. This study aims to develop a facile method of Au nanoparticles (AuNPs) tagged  Riemerella anatipestifer  ( RA ) antisera that can improve these drawbacks. Results demonstrated that citrate-reduction method synthesized AuNPs with an average of 5.0 nm takes a strong extinction band with a maximum absorption peak at 518 nm. 1.0 mM MUA can be coupled onto AuNPs under ultrasonic processing. The carboxylated AuNPs can covalently tagged with  RA  antisera with the activation of EDC/NHS for acquisition of AuNPs/ RA  antisera immunoprobe. Results of DAT and ADID tests affirmed that visualization and sensitivity (at least one concentration gradient) were effectively improved, respectively. The proposed method may offer a new way for large-scale screening of samples.

AuNPs labeled antisera improve the visualization of direct agglutination and sensitivity of agar double immunodiffusion test