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Specific gene and molecular identification method of culicoides cyancus

A specific gene and molecular identification technology, applied in the field of species identification, can solve the problem that there is no gene sequence of Culicoides blue-bellied, and achieve the effect of shortening identification time and ensuring accuracy and reliability

Active Publication Date: 2016-04-13
CHINESE ACAD OF INSPECTION & QUARANTINE
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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 blue-bellied.

Method used

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  • Specific gene and molecular identification method of culicoides cyancus
  • Specific gene and molecular identification method of culicoides cyancus
  • Specific gene and molecular identification method of culicoides cyancus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Acquisition of COI gene sequence of Culicoidescyancus

[0026] 1. Acquisition of specimens of Culicoides blue-bellied

[0027] The specimens of Culicoides blue-bellied were collected in the field, and were directly preserved in 95% alcohol 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 specimen preparation, the end of the thorax and the first few segments of the abdomen of Culicoides blue-bellied were picked and placed in 95% alcohol for DNA extraction.

[0028] 2. DNA template preparation

[0029] The DNA of Culicoides blue-bellied was extracted by the preparation method of micro-DNA templates from small insects (Editor Wen Lizhang. Introduction to Entomological Research Methods and Techniques [M]. Beijing: Science Press, 2010.pp278-286), and the extracted DNA samples were stored in Standby at -20°C.

[0030] 3. Primer synth...

Embodiment 2

[0046] The identification of embodiment 2 unknown Culicoides

[0047] 1. Collection and preservation of specimens

[0048] 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.

[0049] 2. DNA template preparation

[0050] Under a dissecting mirror or a stereo microscope, a female Culicoid was randomly selected from the collected midge specimens, and the Culicoid DNA was extracted using the method of small insect DNA template preparation (Wen Lizhang, editor-in-chief. Entomological research methods and techniques Introduction [M]. Beijing: Science Press, 2010.pp278-286), the extracted DNA samples were stored at -20°C for future use, and each sample had a number.

[0051] 3. Acquisition of target gene sequence

[0052] 3.1 Primer synthesis

[0053]The obtained unknown Culicoides DNA was used as a template to synthesize primers, and the sequences of the primers used were as...

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Abstract

The invention discloses a specific gene and a molecular identification method of culicoides cyancus. The molecular identification method is characterized in that a COI gene sequence of the culicoides cyancus is acquired by adopting a molecular biology method, and species identification is performed on the culicoides cyancus through comparison of the gene sequence similarity. According to the molecular identification method of the culicoides cyancus, achievement of accurate and rapid identification on the culicoides cyancus is facilitated.

Description

technical field [0001] The invention relates to the field of species identification, in particular to a specific gene sequence of Culicoides blue-bellied 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 ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 张晓龙方志强黄恩炯高博滑娜梁慧杰
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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