Real-time fluorescence PCR detection method and kit for sudden death syndrome virus of soybean

A detection kit, soybean sudden death technology, applied in the direction of fluorescence/phosphorescence, biochemical equipment and methods, microbial measurement/inspection, etc., to achieve the effect of short detection time, high sensitivity and strong specificity

Inactive Publication Date: 2014-05-21
SHENZHEN AUDAQUE DATA TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

SDS is mainly distributed in the United States, Argentina, Brazil, Canada, Paraguay, Bolivia, Uruguay and Japan, and there is no report of the disease in China

Method used

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  • Real-time fluorescence PCR detection method and kit for sudden death syndrome virus of soybean
  • Real-time fluorescence PCR detection method and kit for sudden death syndrome virus of soybean
  • Real-time fluorescence PCR detection method and kit for sudden death syndrome virus of soybean

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] 1. Experimental reagents

[0061] Universal Real-time PCR Master Mix was purchased from ABI Company, and DNA extraction kit DNeasy Plant Mini Kit was purchased from QIAGEN Company.

[0062] Primers and probes were synthesized by Shanghai Chaoshi Biotechnology Co., Ltd.

[0063] 2. Experimental equipment

[0064] ABI 7700 real-time fluorescent PCR instrument.

[0065] 3. Experimental method and process

[0066] (1) Design of primers and probes

[0067] First, compare and analyze the genome sequence information of the North American soybean sudden death syndrome pathogen and the South American soybean sudden death syndrome pathogen and their similar species and related species to find out the unique and stable specific base sites of each detection target sequence , the designed alternative primers and probes are shown in Table 1;

[0068] Table 1 Primers and probes for testing

[0069] Numbering

Sequence (5'→3')

Seq ID NO.

FV-F

5′-GGTGCA...

Embodiment 2

[0094] A detection kit is used for real-time fluorescent PCR detection of North American soybean sudden death syndrome pathogen. The kit contains the following primers and probes:

[0095] Primer

[0096] probe

[0097] The kit also includes instructions for use.

Embodiment 3

[0099] A detection kit is used for real-time fluorescent PCR detection of South American soybean sudden death syndrome pathogen. The kit contains the following primers and probes:

[0100] Primer

[0101] The kit also includes instructions for use.

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Abstract

The invention provides a real-time fluorescence PCR method and a kit for detecting sudden death syndrome virus of soybean. By optimizing reaction conditions, the results of the specific testing and sensitivity test show that the detection method has strong specificity and high sensitivity, and is suitable for quick diagnosis and port quarantine for Fusarium tucumaniae and Fusarium virguliforme.

Description

technical field [0001] The invention relates to a detection method and a detection kit for microorganisms, in particular to a real-time fluorescent PCR detection method and a kit for sudden soybean death syndrome (Sudden death syndrome, SDS) bacteria. Background technique [0002] Soybean (Glycine max) is an important oil crop and food crop in my country, and it is also the main source of edible oil and vegetable protein in the world, and occupies an important position in my country's national economy. At present, my country's soybean production ranks fourth in the world, but it still cannot meet the needs of domestic vegetable oil and feed protein processing. As the number of imported soybeans in my country increases year by year, the risk of various diseases that endanger soybean production is gradually increasing. Among them, the two soybean fungal diseases involved in the present invention seriously affect soybean production and trade, and are of economic importance and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/06C12Q1/04C12N15/11G01N21/64
Inventor 章桂明缪建锟王颖程颖慧向才玉
Owner SHENZHEN AUDAQUE DATA TECH
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