Primers and Probes and Reagent Kit for Synchronous Detection of CymMV, ORSV, and CymRSV, and Applications Thereof

A reagent kit for simultaneous detection of CymMV, ORSV, and CymRSV, comprising:

• a set of primers and probes and a real-time fluorescence quantitative PCR amplification system;
• the primers and probe comprising:
• a forward primer CymMV-F, a reverse primer CymMV-R and a probe CymMV-P for CymMV;
• a forward primer ORSV-F, a reverse primer ORSV-R and a probe ORSV-P for ORSV;
• a forward primer CymRSV-F, a reverse primer CymRSV-R and a probe CymRSV-P for CymRSV:
• for CymMV:
• the forward primer, CymMV-F:
• (SEQ ID NO:
• 1)
• 5′-CTGATGCTGGCCACTAACGA-3′;
• the reverse primer, CymMV-R:
• (SEQ ID NO:
• 2)
• 5′-CACGTTCACGGTCAGTAGGG-3′;
• the probe, CymMV-P:
• (SEQ ID NO:
• 3)
• FAM-CCGCCAACTGGGCCAAGGCT-BHQ1;
• for ORSV:
• the forward primer, ORSV-F:
• (SEQ ID NO:
• 4)
• 5′-TTGACCAGTAGGTTCCCTGC-3′;
• the reverse primer, ORSV-R:
• (SEQ ID NO:
• 5)
• 5′-TAGTTGTCGGATTCTGCGGAT-3′;
• the probe, ORSV-P:
• (SEQ ID NO:
• 6)
• HEX-TGGTTACTTCAGAGTTTATCGCTATG-BHQ1;
• for CymRSV:
• the forward primer, CymRSV-F:
• (SEQ ID NO:
• 7)
• 5′-CGCAGTGGGTGACTTATT-3′;
• the reverse primer, CymRSV-R:
• (SEQ ID NO:
• 8
• 5′-CGTCGTGGCTGTGGTAG-3′);
• the probe, CymRSV-P:
• (SEQ ID NO:
• 9)
• Cy5-CACAGTAACCTTCTACGAACCGCAACCG-BHQ3;
• the real-time fluorescence quantitative PCR amplification system comprising:
• 12.5 μL of Premix Ex Taq (2×) mixture and 3.7 μL of RNase-free ddH2O.