Real-time fluorescent RT-PCR detection method, primer and probe for oat mosaic virus and kit

Abstract

The invention relates to the field of molecular diagnosis of biological species and discloses a real-time fluorescent RT-PCR detection method, primer and probe for an oat mosaic virus and a kit. The detection method comprises the following steps: (S1) extracting total RNA of plant tissue from a to-be-detected sample; (S2) introducing a downstream primer OMV-R1 for synthesis of a reverse transcription template cDNA by employing the total RNA as a template; (S3) putting an upstream primer OMV-F1, the downstream primer OMV-R1 and a fluorescent probe OMV-P into a fluorescent quantitative PCR instrument for real-time quantitative fluorescent PCR amplification reaction; and (S4) reading a Ct value to judge the detection result through the fluorescent PCR instrument. A specific primer and a fluorescent probe are designed for a quarantine pest oat mosaic virus; a method for quickly detecting OMV through real-time quantitative fluorescent RT-PCR is built; the method has the advantages of being simple in detection method (closed tube in the whole process and free of electrophoresis detection), high in speed, quantitative, accurate, high in specificity and high in sensitivity, and is suitable for various inspection and quarantine institutions in China for quarantine identification of the quarantine pest oat mosaic virus.

Description

technical field [0001] The invention relates to the field of molecular diagnosis of biological species, in particular to a real-time fluorescent RT-PCR detection method for oat mosaic virus, primers, probes and a kit. Background technique [0002] Oat mosaic virus (OMV) belongs to the Potyviridae family and belongs to the genus Bymovirus. It is linear, often curved, and has no envelope. It has an obvious morphological length, about 600~ 750nm, 12-14nm wide. OMV mainly infects plants of the genus Avena, causing disease of oats sown in autumn, and can cause a 39-60% yield reduction. The spread of OMV mainly depends on the persistent transmission of Polymyxa graminearum, and the long-distance spread is spread with the vegetative material of susceptible plants. At present, OMV is mainly distributed in the United States, Canada, New Zealand, the United Kingdom, France, Ireland and other countries, and has not yet occurred in my country. In 2007, the Ministry of Agriculture's of...

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Problems solved by technology

However, so far, there is a lack of a real-time fluorescent quantitative RT-PCR detection method with strong specificity and high sensitivity for oat mosaic virus

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