A Primer Combination and a Universal Semi-Nested RT-PCR Method for the Simultaneous Identification of Three Porcine Epidemic Diarrhea Virus Strains
A technology for porcine epidemic diarrhea and epidemic diarrhea, which is applied in the field of general-purpose semi-nested RT-PCR and can solve problems such as simultaneous discrimination
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[0027] Simultaneously identify the primer combination of three porcine epidemic diarrhea virus strains, the primer combination is composed of 4 specific primers, which are respectively:
[0028] Upstream primer F1: 5'-ATGGTACGTTGCTAGTGCGTA-3';
[0029] Upstream primer F2: 5'-TGGTGAAATCCAGAGTGTCAAT-3';
[0030] Upstream primer F3: 5'-CCAGCTTATATGCAGGATGGAA-3';
[0031] Downstream primer UR: 5'-TACCATGCACCAAAGTGGAATCAT-3'.
[0032] F1, F2, F3, and UR are respectively shown in sequences 1-4 in the sequence listing.
[0033] The primer design idea of the present invention is as follows image 3 shown.
[0034] The universal semi-nested RT-PCR method for the simultaneous identification of three porcine epidemic diarrhea viruses using the above primer combinations comprises the following steps:
[0035] 1) Extract total RNA from the sample;
[0036] 2) Reverse transcription to obtain sample cDNA;
[0037] 3) Use the above-mentioned primers F1 and UR to carry out the first r...
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