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Specific primers and detection methods for identifying Asian Gypsy Moth

A detection method, technology of gypsy moth, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., can solve problems such as primer sensitivity not meeting requirements and difficulty in distinguishing

Active Publication Date: 2022-03-25
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the egg and larval stages of insects of the genus Gypsy Moth are more difficult to distinguish than the adult stage. When detecting different stages of Gypsy Moth, the sensitivity of some primers cannot meet the requirements, and only adults and late instars can be detected. the larvae
Therefore, there is still a lack of a technology with wide coverage and high enough sensitivity in the rapid identification research of gypsy moth and its relatives.

Method used

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  • Specific primers and detection methods for identifying Asian Gypsy Moth
  • Specific primers and detection methods for identifying Asian Gypsy Moth
  • Specific primers and detection methods for identifying Asian Gypsy Moth

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1 Detection method of Asian gypsy moth

[0039] 1. Extraction of insect genomic DNA

[0040] Genomic DNA of gypsy moth and related species samples collected from different geographical regions in China was extracted by using OMEGA Genomic DNA Mini Kit.

[0041] (1) Crush no more than 50 mg of parasite tissue (pronotum or leg) in liquid nitrogen with a mortar and pestle, and transfer the powder to a clean 1.5 mL microcentrifuge tube.

[0042] (2) Add 350 μl Buffer CTL and 25 μl Proteinase K, vortex and mix well, and incubate at 60°C for 30 min until the sample is completely dissolved.

[0043] (3) Add 350 μl of chloroform:isoamyl alcohol (24:1, v / v) mixture, vortex to mix, centrifuge at 10,000 g for 5 min at room temperature, and carefully transfer the supernatant to a new 1.5 mL centrifuge tube.

[0044] (4) Add an equal volume of Buffer CBL, vortex mix at maximum speed for 15 seconds, and incubate at 60°C for 10 minutes.

[0045] (5) Add an equal volume of a...

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Abstract

The invention discloses a pair of specific primers and a detection method for identifying the Asian gypsy moth. Based on the SS-COI technology, the invention designs a specific region of the mitochondrial COI gene of the Asian gypsy moth for distinguishing other species of the genus Gypsy Primers (SEQ ID NO:1‑2), this pair of primers has accurate identification effects on different geographical populations, different stages and different ages of Asian gypsy moth, and the whole detection process only takes 2‑3 hours, with The advantages of being fast and accurate. It can be used for rapid detection and identification of customs plant quarantine, and provides technical barriers for my country's foreign trade.

Description

technical field [0001] The invention belongs to the technical field of forestry and plant quarantine, and in particular relates to a specific primer for identifying Asian gypsy moth and a detection method. Background technique [0002] Gypsy Moth Lymantria dispar (Linnaeus) is a member of the family Erebidae of the order Lepidoptera, the genus (Lymantria ) is an important quarantine pest. According to its morphological characteristics, female flight ability and geographical distribution area, the latest subspecies classification study divides it into 3 typical subspecies (Pogue & Schaefer, 2007): European subspecies ( Lymantria dispar dispar ), Asian subspecies ( Lymantria dispar asiatica ), Japanese subspecies ( Lymantria dispar japonica ), the European subspecies is also known as European gypsy moth, and the Asian subspecies and Japanese subspecies are collectively referred to as Asian gypsy moth. Compared with the European gypsy moth, the female adults of the Asi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125C12Q1/6844C12Q1/6895
Inventor 石娟季文拽刘姝月
Owner BEIJING FORESTRY UNIVERSITY