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Specific gene of culicoides morisitai and molecular identification method of culicoides morisitai

A specific gene and molecular identification technology, applied in the field of species identification, can solve problems such as the lack of Culicoides Beijing, and achieve the effect of improving identification time and accuracy, and ensuring accuracy and reliability.

Inactive Publication Date: 2018-08-10
南昌市疾病预防控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] There are only a few COI gene sequences of Culicoides in Genbank, but so far there is no related gene sequence of Culicoides Beijing

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Acquisition of Beijing Culicoides morisitai COI Gene Sequence

[0049] 1. Acquisition of specimens of Culicoides Beijing

[0050] The specimens of Culicoides Beijing were collected from mosquito traps in the field. The specimens were killed by freezing and stored in 95% alcohol directly. After the specimens are made into glass slides, they are identified by authoritative experts to ensure the accuracy of the identification results. Before specimen preparation, the thorax and legs of Culicoides pekinensis were picked for DNA extraction.

[0051] 2. Preparation of slide specimens

[0052] Refer to the preparation method of midge specimens (Edited by Yu Yixin, "China Midge", Beijing, Military Medical Science Press, 2006, pp1682-1683)

[0053] 3. Preparation of DNA template

[0054] DNA of Culicoides Beijing was extracted by small insect DNA template preparation method (Wen Lizhang editor-in-chief, "Introduction to Entomological Research Methods and Techniques...

Embodiment 2

[0068] The identification of embodiment 2 unknown Culicoides

[0069] 1. Collection and preservation of specimens

[0070] Lamp lures were adopted on site, and the specimens were harvested in the evening and harvested in the morning. The collected specimens were killed by freezing and stored in 95% ethanol directly.

[0071] 2. Preparation of DNA template

[0072] The DNA of Culicoides Beijing was extracted by using the small insect micro-DNA template preparation method, and the specific method was the same as in Example 1. The extracted DNA samples were stored at -20°C for later use.

[0073] 3. Acquisition of target gene sequence

[0074] 3.1 Primer synthesis

[0075] The obtained unknown Culicoides DNA was used as a template to synthesize primers, and the sequences of the primers used were as follows:

[0076] Forward primer 5'-GGTCAACAAATCATAAAGATATTGG-3'

[0077] Reverse primer 5'-TAAACTTCAGGGTGACCAAAAAAATCA-3'

[0078] 3.2. PCR amplification

[0079] The PCR react...

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Abstract

The invention discloses a specific gene of culicoides morisitai and a molecular identification method of the culicoides morisitai. The method is characterized in that a COI gene sequence of the culicoides morisitai is obtained via a molecular biological method; and through similarity contrast of the gene sequence, species identification of the culicoides morisitai is achieved. According to the molecular identification method of the culicoides morisitai provided by the invention, the accurate and rapid identification of the culicoides morisitai can be achieved.

Description

technical field [0001] The invention relates to the field of species identification, in particular to a specific gene of Culicoides beijing and a molecular identification method thereof. Background technique [0002] Blood-sucking Culicoides are microscopic insects that can transmit and carry pathogens of bluetongue disease, filariasis, Oropouche virus and West Nile disease, and are one of the important medical insects. In recent years, with global warming, the ability of Culicoides to transmit arboviruses has increased to a certain extent, and the distribution range of Culicoides has expanded to the poles and high-altitude areas of the earth. Culicoides populations transmit arboviruses Given the increased capacity of the fauna, some traditionally "non-vector" Culicoides species have the ability to spread disease. The insect-borne diseases transmitted by Culicoides, such as bluetongue disease, deer epidemic hemorrhagic disease, vesicular stomatitis, Schmallenberg disease, e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12Q1/6888C12N15/11
CPCC12N9/0053C12Q1/6869C12Q1/6888C12Y109/03001C12Q2531/113
Inventor 刘仰青陶卉英陈海婴柳小青马红梅
Owner 南昌市疾病预防控制中心