To distinguish the linear from the conformational epitope of SZ34, we used an in-house Western blot/ELISA (Western blot in combination with ELISA) based on polystyrene microspheres. SZ129 (a mAb to VWF A1 domain) was used as a control. Briefly, 20 mg/ml of SZ34 or SZ129 in 0.05 M sodium carbonate buffer (pH 9.6) was mixed with 1:10 volume of 50 mg/ml polystyrene microsphere suspension (SDL37; Takeda Chemical Industries, Co., Ltd., Osaka, Japan). The mixtures were incubated at room temperature for 2 h with shaking. The microspheres were then centrifuged and resuspended in a two-fold dilution of the starting volume of 2.5% (w/v) BSA-PBS to block the free binding sites. This solution was incubated at room temperature for 2 h with constant shaking. After washing three times with PBS, the microspheres were incubated with 100 nM of pVWF with or without denaturing at room temperature for 1 h with gentle agitation. The microspheres were washed three times with PBS, added in sample buffer (0.0625 M Tris-HCl buffer, pH 6.8, 10% (v/v) glycerol, 2% (w/v) SDS, 5% (v/v) b-mercaptoethanol, 0.001% (w/v) bromophenol blue, 4 mM EDTA), and then heated at 99uC for 5 min. After centrifugation at 10,000 g for 3 min, the supernatants were subjected to 6% SDS-PAGE. The proteins were transferred to membranes and blotted with HRP-conjugated anti-VWF IgG (Dako). A Brief Review on Diagnosis of Foot-and-Mouth Disease of Livestock: Conventional to Molecular Tools Foot-and-mouth disease (FMD) is one of the highly contagious diseases of domestic animals. Effective control of this disease needs sensitive, specific, and quick diagnostic tools at each tier of control strategy. In this paper we have outlined various diagnostic approaches from old to new generation in a nutshell. Presently FMD diagnosis is being carried out using techniques such as Virus Isolation (VI), Sandwich-ELISA (S-ELISA), Liquid-Phase Blocking ELISA (LPBE), Multiplex-PCR (m-PCR), and indirect ELISA (DIVA), and real time-PCR can be used for detection of antibody against nonstructural proteins. Nucleotide sequencing for serotyping, microarray as well as recombinant antigen-based detection, biosensor, phage display, and nucleic-acid-based diagnostic are on the way for rapid and specific detection of FMDV. Various pen side tests, namely, lateral flow, RT-LAMP, Immunostrip tests, and so forth. are also developed for detection of the virus in field condition. Foot-and-mouth disease (FMD) is a highly contagious disease affecting artiodactylae, mostly cattle, swine, sheep, goats, and many species of wild ungulates [1] . FMD affects extensive areas worldwide and is included in the list of diseases notifiable to the World Organization for Animal Health (http://www.oie.int/eng/en index.htm). It is recognized as a significant epidemic disease threatening the cattle industry since the sixteenth century and till date it is a major global animal health problem. The history of FMD may be traced to era of Hieronymus Fracastorius, a monk who described a disease outbreak in 1546 A.D. that occurred in cattle near Verona, Italy [2] . Almost 400 years later, in 1897, Friedrich Loeffler and Paul Frosch demonstrated that a filterable agent is responsible for FMD [3] . This was the first demonstration that a disease of animal was caused by a filterable agent and ushered in the era of virology.