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Real-time fluorescent polymerase chain reaction (PCR) detection method for European Lymantria dispar, and primers and probe for detection

A real-time fluorescence and detection method technology, applied in the detection of European gypsy moth, primers and probes for the detection of European gypsy moth, and real-time fluorescent PCR detection of European gypsy moth, which can solve the failure of action and the inability to eradicate completely, etc. problem, to achieve the effect of shortening the detection time

Inactive Publication Date: 2013-07-31
SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In 1890, the U.S. government and state governments began to use spraying to eradicate the gypsy moth. However, the operation failed because it could not be eradicated completely.

Method used

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  • Real-time fluorescent polymerase chain reaction (PCR) detection method for European Lymantria dispar, and primers and probe for detection
  • Real-time fluorescent polymerase chain reaction (PCR) detection method for European Lymantria dispar, and primers and probe for detection
  • Real-time fluorescent polymerase chain reaction (PCR) detection method for European Lymantria dispar, and primers and probe for detection

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Embodiment 1

[0033] 1. Design Primers and Probes

[0034] According to the sequence of some genes in the mtDNA (mitochondrial DNA) of these geographical populations of Gypsy moth, the following specific primers EU-F / R and probe EU-MGB were designed (primers and probes were synthesized by Shanghai Jikang Company):

[0035] European gypsy moth primers:

[0036] Upstream primer EU-F: 5'-AACTTCAGGATGTCCGAAAAATCA-3' (SEQ ID NO.1);

[0037] Downstream primer EU-R: 5'-ACAGCTTTCCTTCTACTTTTTATCTTTACCT-3' (SEQ ID NO.2);

[0038] European gypsy moth probe EU-MGB: FTGGATCTCCCTCCTCCTP (SEQ ID NO. 3) or its complementary chain.

[0039] The detection principle of the above primers and probes is as follows: figure 1 As shown, the alignment of the probe positions of the three different types of gypsy moth sequences is shown in figure 2 .

[0040] 2. Extraction of Gypsy Moth DNA

[0041] Take a single-headed gypsy moth sample (see Table 1 for the source of the sample) and put it into a mortar, add l...

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Abstract

The invention discloses a real-time fluorescent polymerase chain reaction (PCR) detection method for European Lymantria dispar, and primers and a probe for detection. The specific primers EU-F / R and the probe EU-TaqMan are designed according to sequences of partial genes in mtDNA of Lymantria dispar geographical populations such as Asian gypsy moth, European Lymantria dispar and North American Lymantria dispar, and the real-time fluorescent PCR detection method for the European Lymantria dispar is established, so that an aim of quickly and accurately detecting and identifying the European Lymantria dispar is fulfilled.

Description

technical field [0001] The invention relates to the technical field of forestry and plant quarantine, in particular to a detection method for European gypsy moth; in particular to a real-time fluorescent PCR detection method for European gypsy moth; in addition, the present invention also relates to primers for detecting European gypsy moth and probes. Background technique [0002] The gypsy moth Lymantria dispar (Linnaeus) belongs to Lepidoptera, Lyman triidae, and Lymantria, and is recognized worldwide as a pest of forests and ornamental trees in temperate regions of the whole north. Gypsy moth larvae have a wide range of hosts, mainly poplar, willow, elm, oak, larch, linden, spruce and other trees. Mainly distributed in Europe, North America, China, Japan, Korea, according to its morphological characteristics, biological learning and geographical distribution, gypsy moths are divided into Asian subspecies (L. disparasiatica Vnukovsk ii), European subspecies (L. dispardis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 朱雅君林瑶叶军易建平周国梁
Owner SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C