Usage

This test was assessed for sensitivity and application as a rapid diagnostic tool by comparison with virus isolation and detection of PRRS virus antibody in blood. The RT-PCR test was slightly more sensitive than virus isolation for detection of virus in serum and markedly more sensitive than virus isolation from plasma from experimentally infected pigs.

 

Principle

1  RNA in refrigerated serum sample is extracted by TRIzol , The cDNA synthesis was performed with RNA template and specific primer . To produce fragment of PRRSV , degenerate at high temperature, anneal at low temperature, tran script at mid-temperature, cycle with Taq DNA , amplify million fold through 35 times cycle. Electrophoresize RT-PCR products , dyed by ETBR, generate specific fragments in the ultraviolet lights which can be seen by eyes.

 

2  Components:

A      solution: Positive control               60 ul

B      solution: TriZol solution               7ml

C      solution: CHCl₃                      3.5ml

D      solution: C₃H₃O                      4ml

E      solution: CH₃CH₂OH                  12ml

F      solution: DEPC-Deionized water:        500ul

G      solution:RT-PCR buffer               22.5ul X11

H      solution:sample buffer:                50ul

Storage: B, C,D,H, stored at 4-8 ℃;  A,E,F,G, stored at -20 ℃;don’t froze repeatedly.

Expiry date:3 months.

Specification:10 test