Development and Evaluation of a Blocking Lateral Flow Assay Strip for Detection of Newcastle Disease Virus Antibodies

SIMPLE SUMMARY: Newcastle disease (ND) is an Office International Des Epizooties (OIE) notifiable terrestrial and aquatic animal disease that can infect chickens of all ages with a high mortality rate, causing ongoing damage to the poultry industry in China currently. In this study, we have developed a blocking lateral flow assay (bLFA) strip for the rapid detection of Newcastle disease virus (NDV) antibodies, using neutralizing monoclonal antibodies against hemagglutinin-neuraminidase. The results showed that the chicken NDV hyperimmunized serum had a complete blocking titer of 11 log 2, and half-blocking titer of 13 log 2, which are 4 times less than and the same as that of the Hemagglutination inhibition (HI) test (13 log 2), and 8 and 2 times less than that of the virus neutralization test (14 log 2), respectively. Additionally, the bLFA strip has no cross-reactivity with positive serum of other avian pathogens. A total of 510 clinical samples were tested for NDV antibodies. The coincidence rate between the results of the bLFA strip and HI test was 97.65%. Therefore, it is an ideal alternative method for assessing the clinical immunity of ND vaccines in the field in real-time. ABSTRACT: Newcastle disease (ND) is an acute septicemic infectious disease caused by Newcastle disease virus (NDV). Considering that vaccination is currently the main modality for the prevention of ND, it is essential to assess the effectiveness of clinical immunization. In this study, we have developed a blocking lateral flow assay (bLFA) strip for the rapid detection of NDV antibodies using the monoclonal antibody 9C1 against haemagglutinin-neuraminidase (HN), which allows for the determination of an NDV-specific antibody titer within 10 min at room temperature. In addition, the bLFA strip has no cross-reactivity with the positive serum of other avian pathogens including avian influenza subtypes H5, H7, and H9, MD, IBD, IB, EDS, and avian adenovirus. The ability of the bLFA strip for detecting a neutralizing antibody was also estimated. The results showed that the chicken NDV hyperimmunized serum had a complete blocking (100%) titer of 11 log 2, and half-blocking titer of 13 log 2, which are 4 times less than and the same as that of the HI test (13 log 2), and 8 and 2 times less than that of the VN test (14 log 2), respectively. A total of 510 clinical samples were tested for NDV antibodies. The coincidence rate between the results of the bLFA strip and HI test was 97.65%. Therefore, it is an ideal alternative method for assessing the clinical immunity of ND vaccines in the field in real-time.