Abstract
The efficacy of an inactivated foot-and-mouth disease (FMD) vaccine is mainly dependent on the integrity of the foot-and-mouth disease virus (FMDV) particles. At present, the standard method to quantify the active component, the 146S antigen, of FMD vaccines is sucrose density gradient (SDG) analysis. However, this method is highly operator dependent and difficult to automate. In contrast, the enzyme-linked immunosorbent assay (ELISA) is a time-saving technique that provides greater simplicity and sensitivity. To establish a valid method to detect and quantify the 146S antigen of a serotype O FMD vaccine, a double-antibody sandwich (DAS) ELISA was compared with an SDG analysis. The DAS ELISA was highly correlated with the SDG method (R2 = 0.9215, P<0.01). In contrast to the SDG method, the DAS ELISA was rapid, robust, repeatable and highly sensitive, with a minimum quantification limit of 0.06 μg/mL. This method can be used to determine the effective antigen yields in inactivated vaccines and thus represents an alternative for assessing the potency of FMD vaccines in vitro. But it still needs to be prospectively validated by analyzing a new vaccine preparation and determining the proper protective dose followed by an in vivo vaccination-challenge study to confirm the ELISA findings.
Highlights
Foot-and-mouth disease (FMD) is one of the most economically significant trans-boundary diseases among animals; this condition causes severe production losses in domesticated and wild cloven-hoofed animals, in the dairy and pig industries [1]
We abandoned the precipitation step with polyethylene glycol (PEG) 6000 during the purification process because the yields of the 146S antigen were reduced by at least 15% using this method, and a previous study found that PEG precipitation resulted in only a 79.2% recovery of poliovirus infectivity [34]
We developed a double-antibody sandwich (DAS) enzyme-linked immunosorbent assay (ELISA) based on an ELISA and sucrose density gradient (SDG) using nonlinear standard curves and polyclonal antibodies for the quantitative detection of the FMD viruses (FMDV) 146S antigen in live/inactive viral antigen and oil-emulsion vaccines
Summary
Foot-and-mouth disease (FMD) is one of the most economically significant trans-boundary diseases among animals; this condition causes severe production losses in domesticated and wild cloven-hoofed animals, in the dairy and pig industries [1]. Infection with any one serotype does not produce immunity against another serotype. The three most prevalent serotypes in Asia are O, A and Asia 1 [2, 3], while the SAT1 thru SAT-3 serotypes are mainly restricted to sub-Saharan Africa [4]. Vaccination is one of the most practical and effective measures to prevent outbreaks of FMD [5]. Inactivated whole-virus vaccines are produced from cell cultures infected with FMDV and are the most widely used vaccines in China. The use of these vaccines requires strict control of the antigen quality (such as tests for 146S quantification, sterility, identity, purity, safety, potency, stability and immunity) [6,7,8]
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