Methodology Comparison

BoVir^® - real time RT-PCR with Lysis Buffer & amp; Reagents versus ELISA

BoVir® EU and U.S. Protocol

Homogenization 60 minutes at 60º C and 10 minutes
as 95º C

Two-component, one-step RT-PCR master mix

Homogenate pipetted straight from vial to PCR well with no additional RNA extraction necessary

Insert plate into real time PCR instrument

Analyze results

5 Steps
real time RT-PCR – 2.5 hours
Total – 4.0 hours

ELISA EU Protocol

Overnight homogenization of sample

Pipette and incubate 50 µl standard or homogenate for 2 hours at 37°C or overnight at 2-8°C

Washing cycles

Pipette and incubate detection antibody for 1 hour at RT

Washing cycles

Pipette and incubate HRP conjugate for 1 hour at RT

Washing cycles

Pipette and incubate substrate for 30 minutes at RT

Pipette stop solution and read at 450 nm

Analyze results

14 Steps
ELISA – 3.5 hours
Total – 15.5 +/- 2 hours