Narcissus late season yellows virus detection kit and method

A technology for detecting kits and yellowing viruses, which is applied in the direction of biochemical equipment and methods, and the determination/inspection of microorganisms, and can solve the problem of nested RT-PCR technology and molecular detection kits, expensive instruments and equipment for the rapid detection of viruses that have not yet been seen. , port application limitations and other issues, to achieve the effect of avoiding non-specific amplification, high sensitivity, and short detection time
CN103484567BActive Publication Date: 2014-09-24INSPECTION & QUARANTINE TECH CENT OF FUJIAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
INSPECTION & QUARANTINE TECH CENT OF FUJIAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
Publication Date
2014-09-24

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Abstract

The invention relates to a narcissus late season yellows virus detection kit and method specific for narcissus late season yellows virus detection. The knit comprises an outside upstream primer, an outside downstream primer, an inside upstream primer, an inside downstream primer, an RTBuffer, an RNA enzyme inhibiting factor, reverse transcriptase, dNTPs, a PCR Buffer, MgCl2, TaqDNA polymeras, positive control, negative control and RNase-free ddH2O. Specific primers are designed according to narcissus late season yellows virus gene sequences, and a nested RT-PCR technology is utilized to detect narcissus late season yellows viruses. Firstly, inverse transcription is carried out with extracted RNA as the template to synthesize cDNA, first-round PCR amplification is carried out with the cDNA as the template, second-round PCR amplification is then carried out with first-round PCR products as the template, and agarose gel electrophoresis detection is conducted on amplification products: if a specificity target fragment of the size of 539bp appear, the judgment result indicates that a detected virus is a narcissus late season yellows virus. The narcissus late season yellows virus detection kit and method have the advantages of being strong in specificity, high in sensitivity, good in accuracy and short in detection time and are suitable for quickly detecting and monitoring narcissus late season yellows viruses in entry ports, exit ports and agricultural production.
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Description

technical field

[0001] The invention relates to a narcissus late-season yellowing virus detection kit and a detection method thereof, which belong to the field of plant quarantine technology and are suitable for rapid detection and monitoring of narcissus late-season yellowing virus in entry and exit and agricultural production. Background technique

[0002] Narcissus late season yellows virus (NLSYV) belongs to the family Potyviridae (Potyviridae), a member of the genus Potyvirus (Potyvirus), and the virus has similar characteristics to other potato Y viruses. A typical pinwheel-shaped inclusion body will appear in it. The viral genome is a positive-sense single-stranded RNA with a full length of about 9,651bp. Narcissus late-season yellow virus (NLSYV) was first reported in 1980, and since then people have carried out related studies on its virion structure, host range, hazard symptoms, mode of transmission and serological characteristics. The shape of NLSYV granules is ...

Claims

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