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Method for detecting spotted wing drosophila

A Drosophila spotted-winged, amplified fragment technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as inconvenience in quarantine and indistinguishable forms.

Inactive Publication Date: 2012-10-31
中华人民共和国昆山出入境检验检疫局
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During the quarantine work, most of the fruit flies found are larvae or pupae, which are difficult to distinguish from the shape alone, which brings inconvenience to the quarantine

Method used

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  • Method for detecting spotted wing drosophila
  • Method for detecting spotted wing drosophila
  • Method for detecting spotted wing drosophila

Examples

Experimental program
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Embodiment 1

[0017] Embodiment 1 Using the method of the present invention to detect Drosophila spotted wing

[0018] 1 Materials, Reagents and Instruments

[0019] (1) Test material

[0020] See Table 1 for the tested insect samples and related characteristics, and store them in a freezer at -20°C for future use.

[0021] Table 1 Insect samples for testing

[0022]

[0023] UCSD: University of California, San Diego (USA)

[0024] HBU: Hubei University Hubei University

[0025] (2) Instruments

[0026] PCR instrument (Veriti 96, ABI), centrifuge (5415R, Eppendorf), electrophoresis apparatus (EPS300, Tanon), gel imaging system (Gene Genies), vortexer, constant temperature water bath.

[0027] (3) Reagents

[0028] Taq DNA polymerase, dNTP were purchased from Promega, restriction endonuclease Apa I. Hinf Ⅰ and DNA Marker were purchased from Fermentas Company, Agarose Gel DNA Recovery Kit was purchased from Bioengineering (Shanghai) Co., Ltd., primers were synthesized by Nanjing ...

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Abstract

The invention provides a method for detecting spotted wing drosophila, belongs to the technical field of insect quarantine, and particularly relates to a method for detecting spotted wing drosophila. The method comprises the following steps of: extracting a sample genome DNA; amplifying a mitochondrial COI gene sequence and recovering amplification fragments: based on the genomic DNA sample as a template, amplifying the mitochondrial COI gene sequence by adopting COI-F and COI-R, and recovering amplification products; and respectively digesting the amplification fragments by adopting restriction enzyme Apa I and Hinf I, and carrying out agarose gel electrophoresis on digestion products, wherein, two digestion fragments with the lengths of 150-200 and 330-370 are respectively obtained when adopting the restriction enzyme Apa I for digesting the amplification fragments, and the amplification fragments are not cut when adopting the restriction enzyme Hinf I for digesting the amplification products, and at the time, the sample is the spotted wing drosophila. The method is simple, convenient, accurate and sensitive to operate.

Description

technical field [0001] The invention belongs to the technical field of insect quarantine, and in particular relates to a method for detecting spotted winged fruit flies. Background technique [0002] Spotted Drosophila ( Drosophila suzukii Matsumura, 1931) is an important pest on cherries, grapes and other fruits, and there are reports of its occurrence in the United States, Canada, France, Italy, Slovenia, Spain, Switzerland, Japan, Korea and other countries [1] . The fruit fly mainly spreads through host plants and the fruits of host plants. Its damage has attracted widespread attention in the world in recent years. The government departments of the United States, Australia, New Zealand, and EPPO have successively issued early warnings or included it in quarantine. List of pests. [0003] At present, the quarantine and identification of spotted wing Drosophila mainly rely on the relevant morphology of males, while the quarantine and identification of females and other ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 廖太林崔平福吴军袁克纪睿师振华陈集翰孙鹏周锐
Owner 中华人民共和国昆山出入境检验检疫局
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