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Primers, kit and method for detecting potato virus X

A potato and kit technology, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as plant dwarfing, yield reduction, and heavy mosaics and leaves.

Inactive Publication Date: 2018-07-13
陈定虎
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the field, PVX infects potato plants, causing light mosaic or latent symptoms. If infected for many years, it will easily cause heavy mosaic or plant dwarfing, resulting in 10%-80% yield reduction

Method used

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  • Primers, kit and method for detecting potato virus X
  • Primers, kit and method for detecting potato virus X
  • Primers, kit and method for detecting potato virus X

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] A kit for identifying potato virus X RT-LAMP, the kit includes:

[0077]

[0078] Among them, the primer sequence:

[0079] PVX-F3:5'-AGGCCTGTTCACTATCCCG-3'

[0080] PVX-B3:5'-TGATGCCGTTGGAATAGGGA-3'

[0081] PVX-FIP:5'-GCCTCAATCTTGCTGAGGTCCT-AGCCCGTGCCATAGTAGC-3'

[0082] PVX-BIP: 5'-AAGGTGCCCACAGACACTATGG-CTGTTTGAGCGGATGATCCT-3'.

Embodiment 2

[0084] A kit for identifying potato virus X RT-LAMP, the kit includes:

[0085]

[0086] Among them, the primer sequence:

[0087] PVX-F3:5'-AGGCCTGTTCACTATCCCG-3'

[0088] PVX-B3:5'-TGATGCCGTTGGAATAGGGA-3'

[0089] PVX-FIP:5'-GCCTCAATCTTGCTGAGGTCCT-AGCCCGTGCCATAGTAGC-3'

[0090] PVX-BIP: 5'-AAGGTGCCCACAGACACTATGG-CTGTTTGAGCGGATGATCCT-3'.

[0091] The 2× reaction solution buffer described in the present invention is made up of following components:

[0092]

[0093] The enzyme solution is a mixture of Bst DNA polymerase and AMV reverse transcriptase.

[0094] The fluorescent visual detection reagent contains calcein fluorescent dye.

Embodiment 3

[0096] A kind of potato virus X identification method, comprises the following steps:

[0097] A. Plant tissue total DNA extraction;

[0098] B. LAMP detection:

[0099] The kit described in Example 2 was used to configure the pre-reaction solution, and 5 μL of DNA of the sample to be tested was added to the pre-reaction solution to make the total amount reach 25 μL; the control group added 5 μL of deionized water to the pre-reaction solution; positive For the control, add 5 μL of virus-carrying DNA to the pre-reaction solution; use a pipette to mix by suction and drainage, or close the lid and tap lightly to make the solution fully mixed and then centrifuge instantaneously, avoiding bubbles during mixing; place the mixture in the reaction well, React at a constant temperature of 65°C for 90 minutes, and finally keep it at 80°C for 5 minutes to end the reaction;

[0100] C. LAMP result judgment

[0101] After the amplification reaction is over, use an ultraviolet irradiatio...

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PUM

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Abstract

The invention discloses primers, a kit and a method for detecting potato virus X. The primers contain PVX-F3, PVX-B3, PVX-FIP and PVX-BIP. For overcoming the defects in the prior art, the invention aims at providing the primers for RT-LAMP detection of the potato virus X, wherein the primers have the characteristics of fastness, simplicity, convenience, efficiency, high accuracy, high specificityand high sensitivity. The invention also aims at providing the kit which contains the primers and is used for detecting the potato virus X and the method for the RT-LAMP detection of the potato virusX.

Description

technical field [0001] The invention relates to a primer, a kit and a detection method for identifying potato virus X RT-LAMP, belonging to the field of biotechnology. Background technique [0002] Potato virus X, PVX, also known as Potato latent virus or Potato mild mosaic virus, is a model member of Potatovirus X. It is a linear virus composed of a positive strand RNA. About 6.4kbp, the virion is in the shape of a long rod, length×width=515nm×13nm. Under natural conditions, mechanical transmission mainly depends on different parts of the host plant and contact friction between the host plants. The symptoms are mild when PVX is infected alone, and the symptoms are aggravated when combined with Potatovirus Y, PVY, Tobacco veinmottling virus, TVMV, Tobacco etch virus, TEV and other Potatovirus Y viruses , causing serious economic losses. [0003] PVX occurred in all potato Solanum tuberosum production areas, but there were great differences in the infection of different cu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844C12Q2531/119C12Q2563/107C12Q2521/107
Inventor 陈定虎
Owner 陈定虎
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