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Culicoides bellulus specific gene and molecular identification method thereof

A specific gene and gene technology, applied in the field of species identification, can solve the problem of no beautiful Culicoides and achieve the effect of shortening the identification time and ensuring accuracy and reliability

Inactive Publication Date: 2016-05-04
FUJIAN INT TRAVEL HEALTH CARE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, COI gene sequences of various blood-sucking midges have been obtained by this method and uploaded to Genbank, but there is no related gene sequence of Culicoides beautiful

Method used

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  • Culicoides bellulus specific gene and molecular identification method thereof
  • Culicoides bellulus specific gene and molecular identification method thereof
  • Culicoides bellulus specific gene and molecular identification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Acquisition of COI gene sequence of Culicoides bellulus

[0027] 1. Acquisition of Culicoides beautiful specimens

[0028] Specimens of Culicoides glabra were collected in the wild, and were directly preserved in 95% ethanol after being killed by freezing. After the specimens are made into slides, they are identified by authoritative experts to ensure the accuracy of the identification results. Before making specimens, pick the end of the thorax and the first few segments of the abdomen of Culicoides beautiful and place them in 95% alcohol for DNA extraction.

[0029] 2. DNA template preparation

[0030] The DNA of Culicoides beautiful was extracted by the preparation method of small amount of DNA templates from small insects (Editor-in-Chief Wen Lizhang. Introduction to Entomological Research Methods and Techniques [M]. Beijing: Science Press, 2010.278-286), and the extracted DNA samples were stored at -20°C spare.

[0031] 3. Primer synthesis

[0032] T...

Embodiment 2

[0047] The identification of embodiment 2 unknown Culicoides

[0048] 1. Collection and preservation of specimens

[0049] The collected specimens were collected by waving nets or lamp lures, and the collected specimens were killed by freezing and stored in 95% ethanol directly.

[0050] 2. DNA template preparation

[0051] Under a dissecting mirror or a stereo microscope, a female Culicoid was randomly selected from the collected midge specimens, and Culicoid DNA was extracted by using a small-scale insect DNA template preparation method (Wen Lizhang, editor-in-chief. Entomological research methods and techniques Introduction [M]. Beijing: Science Press, 2010.278-286), the extracted DNA samples were stored at -20°C for future use, and each sample was numbered.

[0052] 3. Acquisition of target gene sequence

[0053] 3.1 Primer synthesis

[0054]The obtained unknown Culicoides DNA was used as a template to synthesize primers, and the sequences of the primers used were as f...

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Abstract

The invention discloses a culicoides bellulus specific gene and a molecular identification method thereof. The culicoides bellulus specific gene is characterized in that a molecular biology method is adopted to obtain the COI gene sequence of culicoides, and the species of the culicoides is identified by comparing gene sequence similarity. The ulicoides bellulus molecular identification method is beneficial for accurately and fast identifying culicoides bellulus.

Description

technical field [0001] The invention relates to the field of species identification, in particular to a COI gene sequence of Culicoides and a molecular identification method thereof. Background technique [0002] Culicoides are one of the three major blood-sucking midges in my country. They can transmit bluetongue disease, coccidiosis and other zoonotic diseases, and are an important vector organism. Therefore, the identification and recognition of blood-sucking midges is the basis for monitoring and controlling the occurrence of midge-borne diseases, and it is also a core step in the field of disease prevention and control. At present, the identification of blood-sucking midges mainly depends on the identification of morphological characteristics by experienced taxonomic experts. It is very difficult to identify specimens with related species and incomplete morphological characteristics; moreover, the morphological identification of blood-sucking midges needs to pass The p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/11C12Q1/68
CPCC12N9/0053C12Q1/6869C12Q2531/113C12Q2565/125C12Q2563/173
Inventor 陈敏黄恩炯蔡怡珊高博张建庆王飞鹏
Owner FUJIAN INT TRAVEL HEALTH CARE CENT
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