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Sandwich ELISA

PD Pankaj Deka
SD Sangeeta Das
RH Ritam Hazarika
RK Ray Kayaga
BD Biswajit Dutta
AD Abhijit Deka
UB Utpal Barman
RA Rofique Ahmed
NI Nazrul Islam
MS Mihir Sarma
ID Ilakshy Deka
MR Manoranjan Rout
KS Krishna Sharma
RS Rajeev K. Sharma
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Typing of FMDV in epithelial tissue and vesicular fluid (lesion samples) from suspected infected calves was performed by Sandwich ELISA developed by ICAR-DFMD, Mukteshwar, India. A 10% (W/V) antigen suspension of epithelial tissue was prepared in phosphate-buffered saline (PBS). Briefly, tissue samples were triturated with PBS, and the suspension was mixed with an equal volume of chloroform. After centrifugation at 1000 g for 15 min, the supernatant was collected as antigen and stored at −20 °C for further use. Vesicular fluid was directly used as an antigen for the detection of FMDV.

Type-specific anti-146S FMDV sera raised in rabbits and guinea pigs were used as coating and tracing sera respectively as per the recommendation of ICAR-DFMD, Mukteshwar, India. Rabbit anti-guinea pig IgG-HRP conjugate was used in the assay.

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