Rotavirus Ag

The Rotavirus Ag ELISA is a one-step enzyme immunoassay on the basis of polyclonal antibodies to the group specific VP-6 antigen, the major protein of group A Rotaviruses. Diluted stool specimens and horseradish peroxidase (HRP) labelled polyclonal anti-Rotavirus-antibodies are dispensed simultaneously into the wells of a microtitration plate coated with polyclonal anti-Rotavirus antibodies. After an incubation time of 60 min at room temperature unbound components are removed by a washing step. HRP converts the subsequently added colorless substrate solution of 3,3’,5,5’-Tetramethylbenzidine (TMB) within a 10 min reaction time into a blue product. The enzyme reaction is terminated by sulphuric acid dispensed into the wells turning the solution from blue to yellow. The optical density (OD) of the solution read at 450/620 nm is directly proportional to the specifically bound amount of Rotavirus.