Kit of testing garlic virus and testing method

A detection kit and kit technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of drug waste, complicated operation, test failure, etc., and achieve waste avoidance, high specificity and sensitivity, The effect of easy operation

Inactive Publication Date: 2005-01-12
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no garlic RT-PCR virus detection kit so far. The one-step RT-PCR kits sold on the market carry out the reverse transcription reaction and the PCR reaction in one step. At the same time, the commonly used RT-PCR reaction system is a large system of 100 μl, which causes a lot of waste of medicines, and the operation is complicated and easy to cause pollution, which leads to the failure of the test

Method used

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  • Kit of testing garlic virus and testing method
  • Kit of testing garlic virus and testing method
  • Kit of testing garlic virus and testing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0050] 1. Design and synthesis of PCR primers

[0051] Primers were designed according to the coat protein gene of GMV (Genbank database number: AF500074) and the coat protein gene of GCLV (Genbank database number: AF228416), and 2 pairs of primers GMVP1, GMVP2 and GCLVP1, GCLVP2 were synthesized using a DNA synthesizer. The synthesized primers GMVP1 and GMVP2 amplify the nucleotide sequence located between 277bp and 603bp of the GMV coat protein gene, with a length of 327bp; the synthesized primers GCLVP1 and GCLVP2 amplify the core sequence located between 402bp and 870bp of the GCLV coat protein gene The nucleotide sequence is 469bp in length.

[0052] 2. Assembly of RT-PCR kit

[0053] (1) Assembly of reverse transcription kit

[0054] Reverse transcription reagents were prepared according to Table 1, and distributed in PCR reagent tube 1 (treated with DEPC and sterilized).

[0055] (2) Assembly of PCR kit

[0056] ①PCR reagents were prepared according to Table 2 and d...

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Abstract

This invention relates to garlic virus RT-PCR test reagent box and its test method specially used in the test of GMV and GCLV, among which, the box includes: an inverse transcription reagent box, a PCR box and a positive control. The test method includes reaction of the inverse transcription, PCR reaction and electrophoresis detection, the electrophoresis containing GMV test result is the amplified 32Tbp strip and the result with GCLV is the amplified 469bp strip. The PCR part of the reagent box adopts small reaction system.

Description

1. Technical field [0001] The invention belongs to a garlic virus RT-PCR detection kit and a detection method thereof, and is specially used for the detection of garlic mosaic virus (GMV) and garlic latent virus (GCLV). 2. Technical Background [0002] Garlic (Alliμm sativum L.) is an important export vegetable in my country. However, garlic is a pollen-aborting plant, and it mainly relies on bulbs for asexual reproduction in production. Therefore, the virus continues to accumulate and proliferate in the vegetative organs of garlic, resulting in the degeneration of varieties, the decline of yield and quality, and seriously affecting the production and export of garlic in my country. Garlic virus disease is a disease that is difficult to treat, and there is no effective drug so far. Garlic mosaic virus disease is a disease caused by garlic mosaic virus (GMV, Garlic mosaic virus) infecting garlic plants. The field plant symptoms show obvious yellow stripe mosaic leaves, the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12Q1/25C12Q1/68
Inventor 侯喜林张昌伟史公军王建军
Owner NANJING AGRICULTURAL UNIVERSITY
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