Primers and methods for detecting tobacco mosaic virus lamp

Abstract

The invention discloses a primer and method for detecting tobacco mosaic virus LAMP. The primer comprises a forward outer primer F3:5'-TTACGGAATTACTACTCCATCT-3', a backward outer primer B3:5'-GGTCTAATACTGCGTTGTACC-3', a forward inner primer FIP:5'-TGTTTGGAACTGATTACCTAAGGCA-GTGTTCTTGTCATCAGCGT-3' and a backward inner primer BIP:5'-CTGTCGTTCAACGACAATTCAGC-ACTTTAAAGTCACTATCAGGGA-3'. The method comprises the following steps: extracting tobacco mosaic virus RNA; establishing a LAMP reaction system; adding a pre-reaction solution and a to-be-detected sample into a Loopamp reaction test tube; mixing; putting the mixture into a LAMP turbidity meter for reaction; and detecting the inspection result by a LAMP turbidity meter method or fluorescent visual inspection method. The invention aims at providing a primer and a method for detecting tobacco mosaic virus LAMP, wherein the method has the characteristics of high speed, high detection sensitivity, easiness in operation, direct result reading and the like.

Description

technical field [0001] The invention relates to a primer for detecting tobacco mosaic virus LAMP, and also relates to a method for detecting tobacco mosaic virus by using the primer, which belongs to the field of biotechnology. Background technique [0002] Tobacco mosaic virus, abbreviated as TMV, is an RNA virus that specifically infects plants, especially tobacco and other Solanaceae plants, and is a representative species of the genus Tobamovirus. The virus is Ivanovsky D.I.Iwanowski in 1892 for the first time proved the existence of this TMV. Stanley W.M.Stanley first isolated the TMV virus-like crystals from the squeezed juice of diseased tobacco leaves in 1935. He learned that the protein also contained nucleic acid, and confirmed that the pathogen was the virus. The virus is extremely stable, and because it can proliferate in large quantities in diseased leaves, 2 grams of crystals can be purified from 1 liter of diseased leaf juice. TMV particles are 300 nanometer...

AI Technical Summary

Problems solved by technology

[0006] At present, the detection methods of plant viruses mainly include biology, serology and molecular biology. The biological method is to use the virus indicator plant to detect the presence or absence of the virus, which generally takes 7-10 days; the serological method is based on the antigen-antibody Generally, it only takes about 4-6 hours to detect the virus, but it requires specific antiserum, and the detection sensitivity is not high, and there are still non-specific problems; molecular biology methods mainly refer to the PCR method. The method is fast and has high accuracy, but it requires expensive detection instruments and related equipment, and for viruses like TMV, since its dilution limit is 10 6 times, that is, to dilute the disease juice to 10 6 Therefore, it is necessary to detect the potential TMV with a very high sensitivity detection method. The sensitivity of the above three methods is difficult to achieve, so there will be a possibility of missed detection, so there is an urgent need to study new detection methods

[0007] At present, the existing detection methods have cumbersome detection steps and expensive instruments, and cannot quickly and conveniently identify the authenticity of shark fins.

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