Test system for detection of parainfluenza 3 virus using real time PCR

The test system is designed for detection of parainfluenza 3 virus in blood, tissue or sperm samples from sick or dead animals as well as in infected cell cultures using real time PCR.

The kit contains sufficient reagents to perform 50 tests.

The assay consists of the isolation of the total RNA, reverse transcription reaction and amplification (RT PCR) of the specific fragment using real time PCR.

Real time PCR allows simultaneous amplification and quantification of PCR products.

The amplified fragment of parainfluenza 3 virus DNA can be detected using a molecular beacon (a short chemically synthesised single-stranded oligonucleotide 21 b.p. long) complementary to the inner part of the amplified fragment of the virus that carries a fluorescent label at the 5'-end and a fluorescence quencher at the 3'-end. The beacon undergoes a conformational reorganization in the course of PCR, as the fragment of the virus is amplified and forms a hybrid complex with the beacon, spatially separating the fluorescent label and quencher, thus exposing the fluorescent label. By measuring the fluorescence intensity, the results of PCR amplification can be visualized without gel electrophoresis.

A major advantage is that reaction tubes stay closed during fluorescence measurements, which prevents contamination of the PCR products.

The test system allows differentiation between parainfluenza 3 virus and herpes virus type 1, diarrhoea virus and adenoviruses.

Kit 2 is stored at a temperature between -20 and -18^oC, kit 1 is stored at a temperature between 2 and 6^oC.

12 months.