Kit and detection method for specifically detecting lily cucumber mosaic virus

Abstract

The invention provides an IC-RT-LAMP (Immune Capturing-Reverse Transcription-Loop-Mediated Isothermal Amplification) kit and detection method for specifically detecting a lily cucumber mosaic virus CMV. The kit mainly comprises a specific CMV antibody IgG, a specific RT reverse primer CMV-R, a specific LAMP primer group, a first-chain cDNA (complementary Deoxyribonucleic Acid) synthesis reagent, an LAMP amplification reaction reagent, a fluorescent dye detection solution, a negative reference substance and a positive reference substance; the specific LAMP primer group is composed of a frontward outer primer F3, a backward outer primer B3, a frontward inner primer FIP, a backward inner primer BIP, a positive loop primer LF and a backward loop primer LB; the negative reference substance is ddH2O and the positive reference substance is a Lanzhou lily CMV CP gene standard product. According to the kit provided by the invention, RNA (Ribonucleic Acid) does not need to be extracted and molecular biology special instruments and equipment including an enzyme labeling instrument, a PCR instrument or a fluorescent quantitative PCR instrument and the like are not needed; amplification reaction can be finished in a constant-temperature water bath, and the specificity and sensitivity are high; the kit is convenient to operate and has high applicability; the kit can be used for monitoring generation, spreading and prevalence of the lily CMV and is suitable for being popularized and applied in grassroots.

Description

technical field [0001] The invention relates to an IC-RT-LAMP kit for specifically detecting lily cucumber mosaic virus CMV and a detection method thereof. Background technique [0002] Lily is an important economic crop that integrates ornamental, edible and medicinal uses. Long-term asexual reproduction causes the accumulation of viruses, which seriously affects the yield and quality. Among them, cucumber mosaic virus (CMV) is the most common occurrence. One of the viruses, taking edible lily in Lanzhou as an example, the latest field survey results of viral diseases show that the infection rate of CMV reached 42.5% (Zhang et al., 2018). important factor. [0003] CMV is a typical member of the genus Cucumovirus in the Bromoviridae family. It is distributed worldwide, and its host range is found in dicotyledonous and monocotyledonous plants. As many as 1000 species have been elucidated. More than one species; the molecular weight of CMV virus particles is between 5000 an...

AI Technical Summary

Problems solved by technology

[0010] The technical problem to be solved by the present invention is to overcome the defects and deficiencies in the existing lily CMV virus detection method that require high technical requirements and rely on professional instruments and equipment, and provide a method that can specifically and efficiently detect lily cucumber flowers and leaves without the need for molecular biology professional instruments and equipment. The IC-RT-LAMP method for viruses; specifically, the combination of immunology and LAMP technology is ingenious. Firstly, the specific antibody of CMV virus is coated on the wall of the coated container to specifically capture the CMV particles in the detection sample, and thus To concentrate and purify the virus, and then use the captured CMV particles as a template to perform reverse transcription RT reaction and then LAMP reaction to achieve the purpose of detecting lily cucumber mosaic virus

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