Test system for detection and differentiation of European and American strains of reproductive respiratory syndrome virus using Real-time PCR

The test system is designed for detection and differentiation of European and American strains of porcine reproductive and respiratory syndrome (PRRS) virus in blood, tissue or sperm samples from sick or dead animals as well as in infected cell cultures using Real-time PCR.

The sensitivity of the reaction is 10 to 100 ID₅₀ per ml. 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 PCR, reamplification of specific fragment using Real-time PCR.

The main feature of Real-time PCR - it's possible to register PCR results during the reaction at any given moment of time.

For PCR one pair of universal primers is used (common for both European and American strains of the virus). After RT-PCR fragments 105 b.p. (for European strain) and 96 b.p. (for American strain)are synthesized. Differentiation is done using molecular probes(short one-chained DNA fragments with sizes 21 and 27 nucleotids, chemically synthesized), complementary to internal sites - genomic fragments of PRRSV. The probes have different fluorescent markers at 5’ primers ends. During PCR a probe is destroyed, a marker is released followed by emanation of light. Registering of its intensivity allows to control the process without additional stage - electrophoresis.

Essential advantage of the method is that the registration is done in closed tube, i.e. contamination with PCR products is completely excluded.

The test system allows detection of PRRS of both European and North American types and differentiation between PRRS and other related viruses.

Kit 2 is stored at a temperature between −18 and −20°C, kits 1 and 3 are stored at a temperature between 2 and 6°C.

12 months.