ABSTRACT
Large-scale monitoring of foot-and-mouth disease (FMD) in livestock is imperative in an FMD control program. Detection of antibodies against non-structural proteins (NSP) of FMD virus (FMDV) is one of the best tools to estimate the prevalence of past infection; availability of such a well-validated test is therefore essential. Using a FMDV 3B protein-specific monoclonal antibody, we have developed a new NSP antibody blocking ELISA (10H9 bELISA) and validated it on large panels of sera from different susceptible species. The diagnostic sensitivity of the ELISA was 95% with a specificity of 98%, similar to the values found using a commercial kit (PrioCHECK FMD NS test). The 10H9 bELISA can be used in a broad range of FMD susceptible species making it a very useful tool in monitoring the foot-and-mouth disease control programs by detection of virus circulation in the vaccinated populations. KEY POINTS: ⢠A new ELISA for detection of foot and mouth disease (FMD) antibodies. ⢠Diagnostic sensitivity of 95% and specificity of 98%. ⢠Tested with panels of validated sera from broad host range.
Subject(s)
Foot-and-Mouth Disease Virus , Foot-and-Mouth Disease , Animals , Antibodies, Monoclonal , Antibodies, Viral , Enzyme-Linked Immunosorbent Assay , Foot-and-Mouth Disease/diagnosis , Foot-and-Mouth Disease/prevention & control , Host Specificity , Viral Nonstructural ProteinsABSTRACT
Foot-and-mouth disease virus (FMDV) causes a severe infection in ruminant animals. Here we present an in-depth transcriptional analysis of soft-palate tissue from cattle experimentally infected with FMDV. The differentially expressed genes from two Indian cattle (Bos indicus) breeds (Malnad Gidda and Hallikar) and Holstein Friesian (HF) crossbred calves, highlighted the activation of metabolic processes, mitochondrial functions and significant enrichment of innate antiviral immune response pathways in the indigenous calves. The results of RT-qPCR based validation of 12 genes was in alignment with the transcriptome data. The indigenous calves showing lesser virus load, elicited early neutralizing antibodies and IFN-γ immune responses. This study revealed that induction of potent innate antiviral response and cell mediated immunity in indigenous cattle, especially Malnad Gidda, significantly restricted FMDV replication during acute infection. These data highlighting the molecular processes associated with host-pathogen interactions, could aid in the conception of novel strategies to prevent and control FMDV infection in cattle.
Subject(s)
Cattle Diseases , Foot-and-Mouth Disease Virus , Foot-and-Mouth Disease , Animals , Antiviral Agents/metabolism , Cattle , Cattle Diseases/genetics , Foot-and-Mouth Disease/genetics , Foot-and-Mouth Disease/prevention & control , Foot-and-Mouth Disease Virus/genetics , Foot-and-Mouth Disease Virus/metabolism , Immunity, Cellular , Immunity, Innate/genetics , Viral LoadABSTRACT
Foot-and-mouth disease (FMD) is a major viral disease in farm animals. In the present study, seven monoclonal antibodies (mAbs) were produced against the FMD virus (FMDV)-encoded RNA-dependent RNA polymerase (3D protein) and characterized. Screening of mAb reactivity against three overlapping fragments of the 3D protein expressed in Escherichia coli revealed that the binding sites of all the mAbs were confined to the N-terminal one-third of the 3D protein. A selected mAb was utilized for detecting FMDV in the infected cell culture and tissues obtained from FMDV-infected animals.
