Method for synchronous detection of sweet potato feathery mottle virus (SPFMV), sweet potato virus C (SPVC), sweet potato virus G (SPVG) and sweet potato virus 2 (SPV2)
A technique of synchronous detection and step method, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems affecting the accuracy of detection results, primers cannot distinguish SPFMV and SPVC, short detection cycle, etc., to achieve convenient long-term Time storage, convenient long-distance transportation, and the effect of saving reagent costs
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[0032] Example one:
[0033] 1. Experimental materials
[0034] It was confirmed by a single RT-PCR to be infected with sweet potato G virus (SPVG), sweet potato C virus (SPVC), sweet potato feathery mottle virus (SPFMV), sweet potato virus 2 (SPV2) and multiple infections with SPVG+SPVC, SPVG+SPFMV. , SPVG+SPV2, SPVC+SPFMV, SPVC+SPV2, SPFMV+SPV2, SPVG+SPVC+SPFMV, SPVG+SPVC+SPV2, SPVG+SPFMV+SPV2, SPVC+SPFMV+SPV2 and SPVG+SPVC+SPFMV+SPV2 Sweet potato plants were used as materials, and healthy sweet potato plants were used as negative controls.
[0035] 2. Primer design and synthesis (synthesized by Invitrogen)
[0036] (a) Design and synthesize the universal reverse primer SPFCG2R for detecting SPFMV, SPVC, SPVG and SPV2: 5'-TCGGGACTGAARGAYACGAATTTAA-3', R=A or G, Y=C or T;
[0037] (b) Design and synthesize forward primers SPFF, SPCF, SPGF and SP2F to detect SPFMV, SPVC, SPVG and SPV2 respectively. The primer sequences are as follows:
[0038] SPFF: 5’-GGATTAYGGTGTTGACGACACA-3’,
[0039...
Example Embodiment
[0055] Embodiment two:
[0056] 1. Experimental materials
[0057] The field susceptible sweet potato plants confirmed by a single RT-PCR to be infected with SPVG, SPFMV alone and with SPVC+SPFMV, SPVG+SPFMV+SPV2, SPVC+SPFMV+SPV2 and SPVG+SPVC+SPFMV+SPV2 were used as materials. Healthy sweet potato plants served as negative controls.
[0058] 2. Primer design and synthesis (synthesized by Invitrogen)
[0059] (a) Design and synthesize the universal reverse primer SPFCG2R for detecting SPFMV, SPVC, SPVG and SPV2: 5'-TCGGGACTGAARGAYACGAATTTAA-3', R=A or G, Y=C or T;
[0060] (b) Design and synthesize forward primers SPFF, SPCF, SPGF and SP2F to detect SPFMV, SPVC, SPVG and SPV2 respectively. The primer sequences are as follows:
[0061] SPFF: 5’-GGATTAYGGTGTTGACGACACA-3’,
[0062] SPCF: 5’-GTGAGAAAYCTATGCGCTCTGTT-3’,
[0063] SPGF: 5’-GTATGAAGACTCTCTGACAAATTTTG-3’,
[0064] SP2F: 5’-CGTACATTGAAAAGAGAAACAGGATA-3’,
[0065] Among them, Y=C or T;
[0066] (c) The size of the amplified fragment of...
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