Method for identifying varieties of aedes based on PCR-MS (polymerase chain reaction-mass spectrum) scanning

A PCR-MS, Aedes mosquito technology, applied in the field of genetic engineering, can solve the problems of time-consuming, laborious, dependent on DNA sequencing, etc., and achieve the effect of short time-consuming and high accuracy

Active Publication Date: 2020-06-26
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, morphological identification is the main means of mosquito identification, but it is difficult to identify samples such as incomplete individuals and larvae
DNA barcoding technology can effectively identify the genus and species of mosquitoes, which is not only time-consuming and laborious, but also relies heavily on DNA sequencing

Method used

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  • Method for identifying varieties of aedes based on PCR-MS (polymerase chain reaction-mass spectrum) scanning
  • Method for identifying varieties of aedes based on PCR-MS (polymerase chain reaction-mass spectrum) scanning
  • Method for identifying varieties of aedes based on PCR-MS (polymerase chain reaction-mass spectrum) scanning

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1 establishes gene database

[0043] Establishment of the Aedes gene database: A total of 14,158 related gene sequences were collected through the BOLDSYSTEMS DNA barcode database and NCBI_DNA database, see Table 1 for details.

[0044] Table 1

[0045]

[0046]

Embodiment 2

[0047] Example 2 Establishment of Aedes DNA QR Code

[0048] After clustering analysis with Sequencher5.3 software, select species-specific single nucleotide polymorphism (SNP) as the target, and establish the two-dimensional code type map of Aedes mosquito DNA. For details, see figure 1 .

Embodiment 3

[0049] Embodiment 3 multiplex PCR-MS primer design sequence

[0050] use MassARRAY Assay Designer4.0 software, design multiple PCR-specific primers and probes, see Table 2 for details;

[0051] Table 2

[0052]

[0053]

[0054]

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PUM

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Abstract

The invention discloses a method for identifying varieties of aedes based on PCR-MS (polymerase chain reaction-mass spectrum) scanning. The method comprises the following steps: constructing an aedesvariety identification gene database (COI, COII, 28s, CytB400); constructing an aedes variety identification DNA (deoxyribonucleic acid) two-dimensional code, designing multiple PCR-MS primers, and performing multiple PCR-MS scanning; and performing analysis on thousands of gene sequences of nine aedes varieties, so as to obtain specific SNP (single nucleotide polymorphism) loci of each aedes variety. According to the loci, specific SNP loci of two or more genes of each variety are selected, meanwhile, specific and non-specific loci of other varieties are referred and treated as an exclusive method, analysis software is designed to recognize basic groups of multiple loci, and varieties of aedes can be confirmed, so that the accuracy in identifying aedes varieties can be improved, and detection omission can be avoided.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, and more specifically relates to a method for identifying species of Aedes mosquitoes based on PCR-MS mass spectrometry scanning. Background technique [0002] Mosquitoes can carry and transmit a variety of pathogenic microorganisms. Accurate identification of mosquitoes is of great significance for the prevention and control of mosquito-borne infectious diseases. Aedes mosquitoes such as Aedes albopictus and Aedes aegypti are important vectors of mosquito-borne infectious diseases. Aedes albopictus has a wide distribution range in my country, but with changes in environmental conditions and frequent traffic and trade in various places, the distribution range of Aedes aegypti is gradually expanding. Accurate identification of Aedes albopictus and Aedes aegypti is conducive to better prevention and control of mosquito-borne diseases. [0003] At present, morphological identification ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12Q1/6872
CPCC12Q1/6888C12Q1/686C12Q1/6872C12Q2600/156C12Q2537/143C12Q1/6827C12Q2531/113C12Q2565/627
Inventor 曹晓梅张晓龙杨宇李颖刘莹莹王怀远
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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