Reverse transcription-polymerase chain reaction (RT-PCR) detection for porcine reproductive and respiratory syndrome virus and classical swine fever virus and special primers for same
A technology for RT-PCR and respiratory syndrome, which is applied in the field of RT-PCR detection and its special primers, can solve the problems of high detection cost, time-consuming and laborious, and not fast, and achieve strong sensitivity, good application prospects, and high specificity Effect
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Embodiment 1
[0061] Embodiment 1, multiple RT-PCR detection method construction of porcine reproductive and respiratory syndrome virus, highly pathogenic porcine reproductive and respiratory syndrome virus and classical swine fever virus and design of special primers
[0062] 1. Primer design
[0063] In GenBank, HP-PRRSV (highly pathogenic porcine reproductive and respiratory syndrome virus, Genbank number EF112445), PRRSV (porcine reproductive and respiratory syndrome virus classic strain, Genbank number AY032626), CSFV (swine fever virus, Partially known sequence of Genbank No. AF092448). Homology analysis was performed on each gene region of the virus, and a segment of the NSP2 gene of PRRSV and the E2 gene of CSFV were respectively selected as the target sequence for PCR amplification. Using DNAMAN to design primers for the target sequence, the designed two virus primers (primer pair A can simultaneously amplify the classical strain of porcine reproductive and respiratory syndrome vi...
Embodiment 2
[0101] Embodiment 2, the application of special primer
[0102] 315 clinically suspected disease materials (from the mixed samples of liver, kidney and lung tissues (mass ratio 1:1:1) of dead pigs sent for inspection at the Veterinary Hospital Attached to the Animal Disease Prevention and Control Center of Shandong Province) were tested. as follows:
[0103] Extract 315 RNAs (numbered 1-315) of clinically suspected disease materials, use primer pair A and B as primers, and perform RT-PCR amplification according to the system and procedures in step 2,
[0104] If the RT-PCR product (sequence 5 in the sequence listing) of 525bp is obtained, the sample is positive for PRRSV;
[0105] If the RT-PCR product (sequence 6 in the sequence listing) of 435bp is obtained, the sample is positive for HP-PRRSV;
[0106] If the RT-PCR product (sequence 7 in the sequence listing) of 707bp is obtained, the sample is positive for CSFV;
[0107] If RT-PCR products of 525bp and 435bp are obtain...
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