HDA kit for detecting bacterial leaf streak pathogens of rice and detecting method

A technology of streak bacteria and detection method, which is applied in the field of inspection and quarantine, and achieves the effects of low cost, high sensitivity and accuracy, simple cost and easy operation

Inactive Publication Date: 2015-11-04
JIANGXI NORMAL UNIV
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  • Application Information

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Problems solved by technology

[0006] At present, the research and application of HDA technology in the world is still in its infancy...

Method used

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  • HDA kit for detecting bacterial leaf streak pathogens of rice and detecting method
  • HDA kit for detecting bacterial leaf streak pathogens of rice and detecting method
  • HDA kit for detecting bacterial leaf streak pathogens of rice and detecting method

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Embodiment Construction

[0030] The present invention will be further described in detail below in conjunction with examples. The experimental methods used in the examples of the present invention are conventional methods in the art unless otherwise specified. The reagents or instruments used were not indicated by the manufacturer, and they were all conventional products that could be obtained through commercial channels.

[0031] 1. Design of HDA primer pairs:

[0032] Search the GenBank database for Bacterial leaf spot of oryzae ( Xanthomonas. oryzae pv. oryzicola ) gene sequence and use the BLAST tool in NCBI to perform nucleotide blast comparison to obtain the specific gene of rice bacterial leaf spot pathogen AvrRxo1-ORF1(GenBank number: AY395713.1). The design of the primers is compared with the homology, and the region with high specificity is selected to design the amplification primer pair to increase the accuracy and specificity of the detection. After screening the designed prime...

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Abstract

The invention provides an HDA kit for detecting bacterial leaf streak pathogens of rice and a detecting method. The HDA kit comprises an HDA primer pair for specifically detecting the bacterial leaf streak pathogens of the rice, a 10-fold buffer solution, ATP, dNTPs, Bst polymerase, RecA protein, UvrD helicase, positive control DNA and ddH2O. According to the detecting method, DNA extracted from a sample to be detected is used as a detection template; the bacterial DNA extraction kit is adopted for extracting the DNA of the sample to be detected, and the DNA is conserved for standby; the HDA kit is utilized for carrying out an HDA reaction to obtain an amplified product; the amplified product is detected, and judgment is made. The detecting method has high specificity, can amplify specificity strips only from bacterial leaf streak pathogen DNA of the rice, and has higher sensitivity and accuracy compared with a physiology and biochemistry detecting method; the HDA detecting method can be carried out at constant temperature, and speed is higher compared with that of a conventional PCR detecting method; only a common water bath kettle is needed for carrying out the amplification reaction, and complex and expensive thermal cycler equipment is not needed.

Description

technical field [0001] The invention relates to an HDA kit and a detection method for detecting rice bacterial streak bacterium, belonging to the field of inspection and quarantine. technical background [0002] Rice is one of the most important food crops in the world, and about half of the world's population uses rice as a staple food. Bacterial spot disease of rice, also known as thin strip disease and stripe spot disease, is one of the main diseases of rice, which has seriously affected rice yield. The disease is caused by a bacterium, the pathogen of rice bacterial streak ( Xanthomonas oryzae PV .oryzicola ). In 2007, the Ministry of Agriculture, together with the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, jointly formulated the List of Plant Quarantine Pests Imported by the People's Republic of China, which clearly listed rice bacterial streak disease as a quarantine disease of rice. It is of great s...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 张帆涛谢建坤罗向东周毅
Owner JIANGXI NORMAL UNIV
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