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Detection primer of phakospora sojae sawada, kit and real-time PCR (polymerase chain reaction) detection method

A detection method and technology for detection primers are applied in the field of determination or inspection involving enzymes or microorganisms to achieve the effects of strong specificity, good repeatability and simple detection.

Inactive Publication Date: 2010-08-11
SHENZHEN AUDAQUE DATA TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The commonly used existing polymerase chain reaction (Polymerase Chain Reaction, abbreviated as PCR) molecular diagnostic method can basically meet the accurate and rapid quarantine, detection and identification requirements of some plant pathogenic microorganisms, but EvaGreen dyes are used for real-time PCR and High Resolution Melting (HRM) analysis for the detection of plant pathogenic fungi is still in its infancy at home and abroad

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  • Detection primer of phakospora sojae sawada, kit and real-time PCR (polymerase chain reaction) detection method
  • Detection primer of phakospora sojae sawada, kit and real-time PCR (polymerase chain reaction) detection method

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Embodiment Construction

[0048] The present invention will be further described below in combination with specific embodiments and with reference to the accompanying drawings.

[0049] A real-time PCR detection method for soybean rust bacteria, using detection primers in the detection kit, including forward primer Rs-F and reverse primer Rs-R,

[0050] The sequence of the forward primer Rs-F is:

[0051] SEQ ID NO: 1:PPSG2FP-F:5'-GTGCACTTTTATTGTGGCTCAAAACT-3';

[0052] Has the following nucleic acid sequence:

[0053] PPSG2FP-F: 5'-GTGCACTTTTATTGTGGCTCAAAACT-3' SEQ ID NO: 1;

[0054] The sequence of the reverse primer Rs-R is:

[0055] SEQ ID NO: 2: PPSG2RP-R: 5'-ATGATTAATAGGTGGGTTGCAGC-3'.

[0056] Has the following nucleic acid sequence:

[0057] PPSG2RP-R: 5'-ATGATTAATAGGTGGGTTGCAGC-3' SEQ ID NO:2.

[0058] The specific detection has the following steps in sequence:

[0059]1) Synthetic primers: first design a pair of specific primers consisting of specific forward primer Rsf-F and specific ...

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Abstract

The invention relates to a detection primer of phakospora sojae sawada, a kit and a real-time PCR (polymerase chain reaction) detection method. The primer comprises a forward primer Rs-F and a reverse primer Rs-R, wherein the sequence of the forward primer Rs-F is 5'-GTGCACTTTATTGTGGCTCAAAACT-3', and the sequence of the reverse primer is 5'-ATGATTAATAGGTGGGTTGCAGC-3'. The detection method sequentially comprises the following steps of: synthesizing the primer, adding the primer to an object to be detected and carrying out an amplification reaction, wherein the amplification reaction comprises the following steps of: carrying out pre-denaturation; repeatedly carrying out a reaction interval; and carrying out HRM (High Resolution Melt) analysis on a reaction result automatically recorded by a PCR instrument. The invention has the advantages of simple, quick and accurate detection, strong specificity, high sensitivity, good repeatability and easy popularization and application, can be widely applied to carryING out the bioassay or the detection of phakospora sojae sawada molecules found in the processes of actual quarantines and field surveys and is particularly suitable for occasions having particularly high requirement on timeliness, such as port quarantines, and the like.

Description

technical field [0001] The invention relates to an assay or inspection method comprising enzymes or microorganisms, in particular to a detection primer, a kit and a real-time PCR detection method for soybean rust bacteria. Background technique [0002] Soybean rust (Phakopsora pachyrhizi Sydow), also known as Phakopsora pachyrhizi Sydow, is an obligate parasitic airborne fungus that endangers leguminous economic crops, and belongs to the Basidiomycotina fungus. The host range is currently considered to be limited to leguminous plants. It mainly infects soybean leaves and petioles. In severe cases, stems and pods can also be damaged. It is a phytosanitary dangerous pest prohibited by sovereign countries. It is necessary to establish a rapid detection method for soybean rust, and strengthen port quarantine to effectively prevent soybean rust from entering the country, so as to protect domestic soybean production. [0003] The commonly used existing polymerase chain reaction (...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12R1/645
Inventor 王颖陈枝楠程颖慧章桂明康林仲建忠李芳荣
Owner SHENZHEN AUDAQUE DATA TECH