Detection primers, probe and detection method of highly pathogenic porcine reproductive and respiratory syndrome virus
A porcine blue ear disease virus, highly pathogenic technology, applied in the biological field, can solve the problems of increased detection workload, time and cost, RNA degradation and cross-contamination, and difficulty in realizing high-throughput detection, so as to reduce operation steps and facilitate Degradation, cost-saving effect
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Embodiment 1
[0039] The detection method (direct amplification RT-PCR method) of embodiment 1 highly pathogenic porcine blue ear disease virus
[0040] 1. Design of specific primers and probes
[0041]The highly pathogenic PRRS virus HP-PRRSV has a discontinuous deletion of 30 amino acids in the non-structural protein 2 (NSP2) coding region, based on which specific primers and probes were designed: respectively designed at both ends of the deletion region Upstream and downstream primers; and a Taqman probe was designed to span the flanking sequences at both ends of the amino acid deletion region.
[0042] The sequence of the upstream primer (dHP-PRRSV-F) is:
[0043] 5'-GGTCGGCACCAGTTCCTG-3' (SEQ ID No: 1),
[0044] The sequence of the downstream primer (dHP-PRRSV-R) is:
[0045] 5'-AAATCCAGAGGCTCATCCTGG-3' (SEQ ID No: 2),
[0046] The sequence of the Taqman probe (dHP-PRRSV-pro) is:
[0047] 5'-FAM-CGCGTAGAACTGTGACAACAACGCTGACG-BHQ1-3' (SEQ ID No: 3).
[0048] 2. Exploration of dire...
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