Real time fluorescence reverse transcription polyase chain reaction solid phase reagent kit with one-step method of citrus decline virus and testing method thereof
A decay virus and chain reaction technology, applied in the field of agricultural biology, can solve problems such as the accurate detection of citrus decay virus and the poor specificity of serology kits
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[0055] One, the preparation of citrus decay virus harmless positive control substance: described harmless citrus decay virus positive control substance is made by the following steps:
[0056] (1) extract the total RNA containing citrus decay virus;
[0057] (2) Design the upstream and downstream primers for amplifying citrus decay virus. The upstream primer is characterized by a T7 promoter sequence at the 5' end: 5'-GCAGCTAATACGACTCACTATAGGAACAGACCATGTATAGAGGCGAAACTGCGAAT-3', and the downstream primer sequence is: 5'-TATAGAGGCGAAACTGCGAAT-3' ;
[0058] (3) carry out RT-PCR amplification to obtain the dna fragment of citrus decay virus;
[0059] (4) The RNA fragment of Citrus decay virus was transcribed in vitro by using the DNA fragment of Citrus decay virus as a template, which was used as a harmless positive control.
[0060] 2. The components of the solid-phase detection kit:
[0061] Sample viral RNA extraction reagent, viral RNA fluorescence quantitative reverse tran...
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