Development of immunochromatographic strip assay for rapid detection of novel goose astrovirus

Abstract

The novel goose astrovirus (GoAstV) is an emerging pathogenic virus that has resulted in large economic losses to the goose-rearing industry in China since 2016. The novel goose astrovirus cause gout in goslings with a mortality rate of around 50 %. Therefore, an effective diagnostic approach to monitor the spread of GoAstV is necessary. Here, a novel diagnostic immunochromatographic strip (ICS) assay was developed to detect GoAstV. A rapid immunochromatographic assay based on antibody colloidal gold nanoparticles specific to GoAstV was developed for the detection of GoAstV in goose allantoic fluid and supernatant of tissue homogenate. Monoclonal antibodies (Mabs) were prepared using the hybridoma technology, and the polyclonal antibodies (Pabs) were generated by immunizing the rabbits with recombinant ORF2 protein. In addition, the colloidal gold was prepared by reducing gold salt with sodium citrate coupled with Mabs against GoAstV. The optimal concentrations of the coating antibody and the capture antibody were examined as 1.6 mg/mL and 6 μg/mL. The optimal pH of the colloidal gold labeling was pH 8.0. With the visual observation, the lower limit of the ICS was reported to be approximately 1.2 μg/mL. Common diseases of goose were examined to assess the specificity of the ICS, and no cross-reaction was identified. 40 clinical positive samples were simultaneously detected by using the ICS and the PCR with a 92.5% coincidence rate between them. Furthermore, the mentioned samples could be stored at 25 °C and 4 °C for 4 and 6 months, respectively. It was proved that the ICS in this study was highly specific, sensitive, repeatable and more convenient to rapidly detect GoAstV in clinical samples.