Matched reagent for identifying aedes-aegypti drug resistance and application thereof

A technique for Aedes mosquitoes and reagents, which is applied to a complete set of reagents for identifying drug-resistant traits of Aedes aegypti and its application field

Pending Publication Date: 2017-05-31
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no established molecular detection method and kit for pyrethroid resistance against Aedes aegypti in my country

Method used

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  • Matched reagent for identifying aedes-aegypti drug resistance and application thereof
  • Matched reagent for identifying aedes-aegypti drug resistance and application thereof
  • Matched reagent for identifying aedes-aegypti drug resistance and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1, the preparation of the complete set of reagents of identification Aedes aegypti drug-resistant character

[0065] A complete set of reagents for identifying drug-resistant traits of Aedes aegypti, consisting of primer sets named A and B respectively;

[0066] A is composed of CD1, CD2 and CD3, B is composed of CD1, CD2 and CD4, CD1 is the single-stranded DNA shown in SEQ ID No.1, CD2 is the single-stranded DNA shown in SEQ ID No.2, and CD3 is SEQ ID The single-stranded DNA shown in No.3, CD4 is the single-stranded DNA shown in SEQ ID No.4.

[0067] CD1 and CD2 can amplify the 587bp DNA fragment shown in sequence 5 from the wild-type Aedes aegypti genome or the Aedes genome containing the V1016G mutation, and CD2 and CD3 can be amplified from the wild-type Aedes aegypti genome The 344bp DNA fragment shown in sequence 6, CD2 and CD4 can amplify the 344bp DNA fragment shown in sequence 7 from the Aedes aegypti genome containing the V1016G mutation.

Embodiment 2

[0068] Embodiment 2, Aedes aegypti drug resistance detection

[0069] Genomic DNA of 100 Aedes aegypti mosquitoes numbered AA001-AA100 captured in the field were extracted respectively.

[0070] The genomic DNA of each Aedes aegypti was subjected to PCR amplification according to the following method (amplification system is shown in Table 1):

[0071] (1) Take reagent T (Treasure Bioengineering Co., Ltd., product number DR011, the components of reagent T are shown in Table 2) and add them to 2 PCR tubes, labeled tube A and tube B, 12.5 μL in each tube.

[0072] (2) Add 1.1 μL of reagent P1 to tube A, and add 1.1 μL of reagent P2 to tube B. Reagent P1 is the solution that CD1, CD2 and CD3 of embodiment 1 and water form, and the molar ratio of CD1, CD2 and CD3 is 3:4:4 in the reagent P1; Reagent P1 is the CD1 of embodiment 1, CD2 and CD4 and water The solution composed, the molar ratio of CD1, CD2 and CD4 in reagent P1 is 3:4:4.

[0073] (3) Add 9.9 μL deionized water to tub...

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PUM

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Abstract

The invention discloses a matched reagent for identifying the aedes-aegypti drug resistance and an application thereof. The matched reagent for identifying the aedes-aegypti drug resistance is prepared from a primer set A and a primer set B; the A is prepared from CD1, CD2 and CD3, and the B is prepared from CD1, CD2 and CD4; the CD1 is single stranded DNA shown in the SEQ ID No.1, the CD2 is single stranded DNA shown in the SEQ ID No.2, the CD3 is single stranded DNA shown in the SEQ ID No.3, and the CD4 is single stranded DNA shown in the SEQ ID No.4. The matched reagent for identifying or identifying aedes aegypti in an auxiliary mode can be used for identifying the pyrethroid drug resistance condition of the aedes aegypti, and is fast and convenient, simple and convenient and sensitive, and it is identified that the accurate rate of the pyrethroid drug resistance condition of the aedes aegypti is 100%.

Description

technical field [0001] The invention relates to a set of reagents for identifying drug-resistant traits of Aedes aegypti in the field of biotechnology and an application thereof. Background technique [0002] Aedes aegypti (Aedes aegypti) is an important species of blood-sucking mosquito in tropical areas, and is the main vector of various infectious diseases such as dengue fever, dengue hemorrhagic fever, yellow fever, and chikungunya fever. In recent years, dengue fever epidemics have occurred successively in Guangdong, Yunnan and other places, and the number of reported cases has continued to rise, making the epidemic situation quite severe. Dengue fever is one of the most important vector vector diseases with the widest distribution, the largest number of cases and the greatest harm in the world. Since the disease has no vaccine prevention and specific drug treatment so far, vector control measures have become the most effective way to prevent and treat the disease. In...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/106C12Q2600/156
Inventor 李春晓赵彤言董言德郭晓霞张恒端张映梅邢丹
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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