Aedes albopictus molecular marker related to deltamethrin insecticide resistance, primer and application

A molecular marker, Aedes albopictus technology, applied in the field of molecular biology, can solve the problem of no mosquito function verification report and other problems

Pending Publication Date: 2021-10-22
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the direct relationship between the non-synonymous mutation of the mosquito VGSC gene and the resistance to pyrethroid insecticides has not been reported in the mosquito body.

Method used

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  • Aedes albopictus molecular marker related to deltamethrin insecticide resistance, primer and application
  • Aedes albopictus molecular marker related to deltamethrin insecticide resistance, primer and application
  • Aedes albopictus molecular marker related to deltamethrin insecticide resistance, primer and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Screening of laboratory Aedes albopictus deltamethrin-resistant populations

[0045] The sensitive strain of Aedes albopictus in the laboratory comes from the sensitive strain domesticated and bred by Shanghai Center for Disease Control and Prevention. The offspring hatched by laying eggs of this sensitive population is used as the first generation population, and each generation is screened. Before each generation of screening, at first measure the median lethal concentration of contemporary larvae (see table 1 in embodiment 2), then add in the special-purpose steel basin of screening with the deltamethrin liquid medicine of the corresponding concentration that is configured into 1L volume with contemporary median lethal concentration, Then add about 1000 third-instar Aedes albopictus larvae for screening, and after 24 hours, suck out the surviving larvae with a straw and transfer them to dechlorinated tap water for further feeding and passage. Use 2 / 3 of th...

Embodiment 2

[0046] Embodiment 2 Aedes albopictus larvae resistance detection

[0047] The larvae were selected by the larva dipping method recommended by the World Health Organization for the determination of larval drug resistance, and the half lethal concentration (LC) of the test population was obtained. 50 ).

[0048] First, accurately weigh the deltamethrin standard (94.62%, provided by the Chinese Center for Disease Control and Prevention), and use acetone as a solvent to prepare a mother solution and dilute to obtain 1 mL of the drug solution with different concentrations for use. Mix 25 3-4 instar Aedes albopictus larvae with 99mL dechlorinated tap water and different concentrations of liquid medicine into a disposable paper cup as the experimental group, and the control group is made by mixing 99mL dechlorinated tap water and 1mL acetone. Three replicates were set up for each test concentration and control group. The test is carried out in the pre-experiment first, and generall...

Embodiment 3

[0055] Example 3 Detection of Aedes albopictus Adult Mosquito Resistance

[0056] Adult mosquitoes use the WHO adult mosquito contact cylinder drug-film filter paper contact method. The standard bioassay cylinder designated by the World Health Organization was assembled, and 0.05% deltamethrin film and control film were respectively loaded into the contact cylinder. There were five experimental groups and two control groups in each experiment. Use a mosquito suction tube to take 20-25 female Aedes albopictus 3-5 days after eclosion and not sucking blood into the recovery cylinder, observe for 15 minutes and record whether any adult mosquitoes are dead or damaged. After 15 minutes, the mosquitoes in the recovery cylinder were blown into the contact cylinder, and contacted with the drug film for 1 hour, during which the number of adult mosquitoes knocked down was recorded every 10 minutes. After 1 hour, the adult mosquitoes in the contact cylinder were transferred back to the c...

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Abstract

The invention discloses an aedes albopictus molecular marker related to deltamethrin insecticide resistance. The locus of the molecular marker is located at the 4421st site of a VGSC gene cDNA sequence shown as SEQ ID NO: 1, the locus of the molecular marker has T->C mutation, amino acid mutates from phenylalanine (F) to serine (S), and the mutation is at the 1474th site of a VGSC gene amino acid sequence shown as SEQ ID NO: 2. The invention also discloses a primer for amplifying the molecular marker and a method for detecting the molecular marker by using the primer. The invention also discloses an application of the molecular marker in on-site detection of the resistance of a mediator insecticide deltamethrin. The method overcomes the defects of a traditional adult mosquito bioassay method, has an application prospect of serving as a sensitive, specific and easy-to-operate medium field population resistance monitoring method, and has a potential value in the aspect of mosquito-borne infectious disease prevention and control.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to molecular markers, primers and applications of Aedes albopictus related to deltamethrin insecticide resistance. Background technique [0002] Dengue fever is a mosquito-borne viral disease caused by dengue virus (DENV). It is prevalent in 128 countries around the world. There are 390 million infected patients every year, and about 2.5 billion people are at risk of contracting dengue fever. Most prevalent in developing countries, the WHO lists it as one of the top ten global health threats. Dengue fever is the most important mosquito-borne infectious disease in my country at present, and it has been popular to varying degrees every year in the past 40 years. In 2014, dengue fever was prevalent in almost all prefecture-level cities in Guangdong, with more than 37,000 confirmed cases of dengue fever. [0003] Aedes albopictus is the main vector of dengue fever...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/156C12Q2600/124C12Q2537/143C12Q2521/327
Inventor 郭怡佳陈晓光
Owner SOUTHERN MEDICAL UNIVERSITY
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