Primers for detecting plum pox viruses as well as kit and detection method

Abstract

The invention discloses primers for detecting plum pox viruses as well as a kit and a detection method, wherein the primers comprise the forward direction outer primer F3: 5'-GATGATGGATGGGGAAACA-3', the reverse direction outer primer B3: 5'-AGGCTGTAGTCTGTCAGG-3', the forward direction inner primer FIP: 5'-GCCATAATTTGTCTAAAAGTGGGTT-CAAGTGGAGTATCCAATAAAGC-3', and the reverse direction inner primer BIP: 5'-ACATTTCAGTAACGTGGCTGAA-GCTGAATCCCATACCTTGG-3'; the kit comprises the primers and the detection method adopts the primers; with adoption of the primers and the detection method, the plum pox virus detection is high in speed, simple and convenient, efficient, high in accuracy, strong in specificity and high in sensitivity.

Description

technical field [0001] The invention relates to a primer, a kit and a detection method for detecting plum pox virus, belonging to the field of biotechnology. Background technique [0002] Plum pox virus (PPV) is one of the most dangerous viruses for stone fruit trees. Once the fruit tree is infected by it, it will cause a large number of unripe fruits to fall off, severely reduce the yield, and greatly reduce the quality of the fruit. , and the virus spreads fast, which may bring catastrophic losses to the development of the fruit tree industry of the whole country in a short period of time. The virus was first discovered in Bulgaria in 1915, and then quickly spread to most countries in Europe, as well as the Mediterranean, Central Africa, India and Chile, and has also been reported in the United States and Canada. Because of the seriousness of the virus, it has been listed as a quarantine pathogen of type A2 by the European Plant Protection Organization. A series of stric...

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Problems solved by technology

However, high-efficiency antiserum must be available for serological reactions. In addition, serological testing cannot accurately measure some samples with low virus content and interfering substances, and false positive results are prone to occur, and there is a problem of long detection time. The problem is that it cannot meet the requirements of rapid detection at the current port; RT-PCR technology is one of the most widely used new technologies in molecular biology. Compared with ELISA detection, the detection of viruses by RT-PCR technology has the characteristics of high sensitivity and can be detected. Minimum virus content to reach fg level

At present, many new technologies are continuously derived on the basis of RT-PCR technology, such as real-time fluorescent RT-PCR technology, gene chip technology, etc., so that the scope of application is continuously expanded, and the specificity and sensitivity are also continuously improved. However, these molecular biological However, RNA extraction is cumbersome and easy to degrade, and PCR reaction is easily interfered by polyphenols and polysaccharides in the host, which limits the rapid, simple and efficient detection of plant virus disease samples

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