oligo nucleotide PRIMERS, FLUORESCENT PROBE AND METHOD FOR IDENTIFYING RNA OF THE VIRUS OF DISEASE IBARAKI BY POLYMERASE CHAIN REACTION B THE REAL-TIME METHOD

摘要

1. Synthetic oligonucleotide primers and a probe complementary to conserved region of the genome segment 10 Ibaraki disease virus, used to identify the genome fragment VBI and having the following base composition: IbarF 5'-GATCAAACCATTTTGCGCTT-3'IbarR 5'-CTCATCCTCACCGCCTCATTG-3'IbarZ 5'- [HEX] TCTTGTATGGTCAATCCGCTGGCT [BH2] -3'.2. A method of detecting Ibaraki disease virus, comprising isolating RNA from biological material, setting a reverse transcription reaction for cDNA synthesis, formulation polymerase chain reaction in real time using the designed primers and probe of claim 1, the amplification of viral RNA, evaluation of the reaction, wherein that the synthesis of the cDNA, which in turn consists of RNA chains denaturation primers and IbarF IbarR for 10 min at 88 ° C and cDNA synthesis on template RNA VBI for 30 min at 42 ° C, then n oestriasis keeping the reaction results through the unit's software, the result of the reaction is considered positive when fluorescence curve analysis sample intersects a threshold line.