Method for rapidly identifying opogona sacchari (entry plant quarantine pest)

A technology for plant quarantine and harmful organisms, which is applied in the biological field and can solve cumbersome problems

Inactive Publication Date: 2010-07-28
HUNAN AGRICULTURAL UNIV +1
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  • Abstract
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Problems solved by technology

[0003] At present, the common method of detection and identification of sucrose moth in my country is still the laboratory morphological identification ...

Method used

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  • Method for rapidly identifying opogona sacchari (entry plant quarantine pest)
  • Method for rapidly identifying opogona sacchari (entry plant quarantine pest)
  • Method for rapidly identifying opogona sacchari (entry plant quarantine pest)

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Embodiment Construction

[0113] Taking the organisms on the plants to be inspected that are about to enter the country as samples, the extraction of the sample genomic DNA, the concentration and purity determination of the genomic DNA extract, the PCR reaction system ( The total reaction system is 25 μl, and the components are as follows: 2.5 μl of 10×PCR Buffer, 2 μl of each 2.5 mM dNTP Mixture, 1 μl of each 10 pM sucrose moth-specific primer CF720 / CF1113, 0.13 μl of 5 U / μl Taq DNA polymerase, 50 ng / μl DNA extraction solution 2 μl, sterilized double distilled water 16.37 μl) and the set PCR reaction parameters (pre-denaturation at 94°C for 4 min; denaturation at 94°C for 20 s, annealing at 62°C for 20 s, extension at 72°C for 30 s, a total of 30 cycles; final 72°C Extend for 5 minutes, store at 4°C) for PCR amplification; after the specific amplification is completed, take 10 μl of PCR amplification solution and 2 μl of 6x sample buffer to mix, and the mixture is placed in 1.5% agarose gel at a volta...

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Abstract

The invention discloses a method for rapidly identifying opogona sacchari (an entry plant quarantine pest), which comprises: performing PCR amplification of the extract of the genome DNA of a sample to be tested by using opogona sacchari specific primer CF720/CR1113; performing electrophoresis on PCR amplification solution in 0.5XTE electrophoresis buffer under a voltage of 5V per centimeter of gel; and placing the PCR amplification solution in an ethidium bromide staining box for staining, and observing an amplification band with an ultraviolet gel imaging device to determine if the tested is opogona sacchari (the entry plant quarantine pest). The method has the advantages that: the identification time is reduced to less than 24 hours from the original more than one week; and the identification accuracy is improved to 99 percent from the original 70 percent. The method can help entry and exit quarantine and inspection department, customs and other related units to perform quick, accurate and high efficiency detection and identification at ports.

Description

Technical field: [0001] The invention relates to the field of biotechnology, in particular to a rapid identification method for the imported plant quarantine pest sucrose moth. Background technique: [0002] Opogona sacchari (Bojer, 1856) is a phytosanitary pest imported into my country (see the 2007 edition of "The List of Phytosanitary Pests Imported by the People's Republic of China"). It mainly harms ornamental garden plants such as Brazilian wood, rich tree and Palmaceae, as well as traditional economic crops such as bananas and sugar cane. For example, in Beijing, the annual elimination rate of Brazilian wood in severely damaged greenhouses is over 50%, causing economic losses of up to several million yuan. [0003] At present, the common method of detection and identification of sucrose moth in my country is still the laboratory morphological identification method, which requires cumbersome steps such as rearing, dissection, and microscopic examination, and the ident...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 黄国华王星廖力徐淼锋盛夏冰李杰
Owner HUNAN AGRICULTURAL UNIV
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