Development and Validation of a Competitive ELISA Based on Bovine Monoclonal Antibodies for the Detection of Neutralizing Antibodies against Foot-and-Mouth Disease Virus Serotype A

Abstract

The level of neutralizing antibodies in vaccinated animals is directly related to their level of protection against a virus challenge. The virus neutralization test (VNT) is a “gold standard” method for detecting neutralizing antibodies against foot-and-mouth disease virus (FMDV). However, VNT requires high-containment facilities that can handle live viruses and is not suitable for large-scale serological surveillance. In this study, a bovine broadly neutralizing monoclonal antibody (W145) against FMDV serotype A was successfully produced using fluorescence-based single-B-cell antibody technology.

Optimization of a foot-and-mouth disease virus Southern African Territories–specific solid-phase competitive ELISA for small ruminant serum samples

“…Similar findings were reported for serotype O experimentally infected sheep 26 and cattle, 21 as well as cattle and swine vaccinated with serotype A. 5 Although an initial serum dilution of 1:10 is employed when testing cattle sera, the results of a previous study suggested that a higher serum dilution might be required for goat sera. 9 We selected the 1:15 dilution as optimal because the agreement was higher, the Youden index was greater at the selected cutoff (balance between the DSp and DSe was most favorable), and the optimal cutoff was closest to the typically recommended value of 50% (middle of the expected linear range of the test).…”

Development of gold nanoparticles-lateral flow test as a novel field diagnostic assay for detecting foot-and-mouth disease and lumpy skin disease viruses

“…The difference in the obtained values of the particle size of the Au-NPs is based on the difference between the observable particle edges, mono-disperse could be owing to the fact that the existence of some significant bio-organic molecules in the plant extract seems to act as a legend which efficiently stabilizes the formed gold nanoparticles and responsible for reducing the Au+ to Auº as shown in Fig 1 , and this explanation was agreed with ( 35 ). The protein conjugated with an antibody, was found to be more stable as shown in Fig 1 .…”

Development and Evaluation of a Monoclonal Antibody-Based Blocking Enzyme-Linked Immunosorbent Assay for the Detection of Antibodies against Novel Duck Reovirus in Waterfowl Species

“…However, there are some limitations, including that it uses live virus stocks, it is time-consuming and laborious, it has poor repeatability, and it is not suitable for assaying a large number of samples at one time, which limits the routine applicability of the VNT ( 37 – 42 ). Moreover, although the conventional ELISA is simple and rapid ( 20 ), it is unsuitable for large-scale screening of NDRV disease in waterfowl species, mainly due to the fact that the routine ELISA requires species-specific conjugates for the detection of NDRV antibodies in waterfowl species, which are currently unavailable ( 35 , 43 ).…”

“…Currently, the routine detection methods for the NDRV pathogen and associated antibodies include virus isolation ( 4 , 10 , 14 ), RT-PCR ( 17 , 18 ), virus neutralization test (VNT) ( 13 ), and indirect enzyme-linked immunosorbent assay (ELISA) ( 19 ). ELISA is a powerful technique for detecting and measuring antigens and antibodies ( 20 ). This method has advantages such as ease of operation and high sensitivity and specificity, making it much more suitable for assessing immune responses and epidemiological surveys.…”