Preparation of monoclonal antibodies against norovirus and establishment of a rapid immunochromatographic technique

Abstract

Norovirus is one of the main pathogens causing acute gastroenteritis worldwide, and possesses rich genetic diversity. We established a simple, fast, and suitable colloidal gold immunochromatographic detection method for common epidemic strains of Norovirus. A pair of monoclonal antibodies that are sensitive to recognizing Norovirus was successfully screened and used as gold-labeled antibody and capture antibody, respectively. The assembly conditions of the test strip were optimized to determine the labeling pH, the concentration of capture antibody and concentration of AuNP-labeled antibody, then the performance of the strips, including sensitivity testing and specificity testing, was evaluated. The minimum detection concentration of the established Norovirus colloidal gold test strip detection method is 10 ng mL⁻¹. This antibody has no cross-reactivity with HCoV-NL63, HCoV-OC43, Influenza A (H1N1), Influenza A (H9N2), respiratory syncytial virus or Varicella zoster virus. The established colloidal gold detection method was used to analyze 11 fecal samples, and the results showed that only two samples were positive and the rest were negative, in agreement with the RT-PCR result. The whole colloidal gold test strip detection method process took 15 min, and could be used for on-site detection. The established colloidal gold test strip detection method has good specificity and stability, and can be used for the detection of common Norovirus strains and in large-scale epidemiological investigations.