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Specific primer, kit and method used for detecting chrysomyxa

A specificity and kit technology, applied in the field of specific primers and detection for the detection of golden rust bacteria, can solve problems such as identification, and achieve the effects of high accuracy, good primer specificity and remarkable detection effect.

Inactive Publication Date: 2015-09-23
BEIJING FORESTRY UNIVERSITY
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Problems solved by technology

However, conventional disease diagnosis techniques are difficult to quickly and accurately identify the pathogenic bacteria of the disease

Method used

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  • Specific primer, kit and method used for detecting chrysomyxa
  • Specific primer, kit and method used for detecting chrysomyxa
  • Specific primer, kit and method used for detecting chrysomyxa

Examples

Experimental program
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Effect test

Embodiment

[0041] 1. Primer design and synthesis

[0042]Use software such as Clustal X and MEGA to carry out base difference comparison analysis on the ribosomal gene ITS region sequence of the known type strain of Aureus aureus, design specific primers according to the analysis results, and obtain the sequence table as SEQ ID No.1 and SEQ ID The primer pair shown in No.2, its base sequence is respectively:

[0043] F: 5'-CCTGCGGAAGGATCATTATT-3'

[0044] R: 5'-TAAGTTCAGCGGGTAGTCCC-3'.

[0045] The above primers were analyzed by BLAST in GeneBank to verify their specificity, and entrusted to Shenzhen BGI Co., Ltd. to synthesize them to obtain the above primers.

[0046] 2. Preparation of DNA template

[0047] Pick 100-200 rust spores from a single pile of rust spores on spruce needles under a dissecting microscope, place them on a sterilized glass slide, and mark the concentrated area of ​​rust spores as an "O" shape , gently cover another piece of sterilized glass slide while pressi...

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PUM

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Abstract

The invention discloses a specific primer used for detecting chrysomyxa. The primer is composed of a forward primer body and a reverse primer body. The nucleotide sequence of the forward primer body is CCTGCGGAAGGATCATTATT. The nucleotide sequence of the reverse primer body is TAAGTTCAGCGGGTAGTCCC. The invention further provides a kit based on the primer and a method for detecting the chrysomyxa through the primer. By means of the specific primer, kit and method, accuracy is high, sensitivity is high, whether the chrysomyxa exists in samples or not can be rapidly and easily judged, and an effective means can be provided for disease early-stage monitoring.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a specific primer and a detection method for detecting aureus. Background technique [0002] Chrysomyxa Unger mainly infects Pinaceae plants such as Picea, Tsuga and Keteleeria, causing damage to needles, cones and terminal buds, causing spruce Serious diseases such as leaf rust, spruce cone rust and spruce bud rust. Widely distributed in Sichuan, Gansu, Shaanxi, Xinjiang, Northeast China and Qinghai, etc., it causes the decline of spruce forests, threatens the safety of natural and artificial spruce forests, and causes huge economic losses. In the latest "National Forestry Dangerous Pest List" published by the State Forestry Administration in January 2013, the 36 important tree pathogenic fungi include three important species of aureus that cause spruce leaf rust and cone rust: Chrysomyxadeformans (Diet.) Jacz., Qinghai spruce leaf rust (ChrysomyxaqilianensisWang.Wu et Li) ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2531/113
Inventor 游崇娟田呈明梁英梅
Owner BEIJING FORESTRY UNIVERSITY
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