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Kit for detecting drug resistance of culex pipiens and special primer thereof

A technology of Culex pipiens and kit, applied in the direction of DNA/RNA fragment, recombinant DNA technology, etc.

Inactive Publication Date: 2011-11-16
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

Although AS-PCR technology has been applied to the detection of mosquito resistance, most of them are established for a single point mutation of the kdr gene
At present, there is no molecular detection method and kit that can detect multiple mutation sites at the same time

Method used

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  • Kit for detecting drug resistance of culex pipiens and special primer thereof
  • Kit for detecting drug resistance of culex pipiens and special primer thereof
  • Kit for detecting drug resistance of culex pipiens and special primer thereof

Examples

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Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1, detection of drug resistance of Culex apices

[0068] 1. PCR amplification

[0069] 1. Design of primers

[0070] Specific primers were designed for the kdr (Accession number: AY573288) gene associated with resistance to pyrethrins in Culex aceritoides:

[0071] CD1: 5'AAC TTC ACC GAC TTC ATG CAC 3' (SEQ ID NO: 1, downstream primer)

[0072] CD2: 5'CAA GGC TAA GAA AAG GTT AAG AAC 3' (SEQ ID NO: 2, upstream primer)

[0073] CD3: 5'CCA CCG TAG TGA TAG GAA ATT TA 3' (SEQ ID NO: 3, downstream primer)

[0074] The specific primer designed according to the mutant kdr L1014F type mutant gene (sequence 6) of kdr:

[0075] CD1: 5'AAC TTC ACC GAC TTC ATG CAC 3' (SEQ ID NO: 1, downstream primer)

[0076] CD2: 5'CAA GGC TAA GAA AAG GTT AAG AAC 3' (SEQ ID NO: 2, upstream primer)

[0077] CD4: 5'CCA CCG TAG TGA TAG GAA ATT TA 3' (SEQ ID NO: 4, downstream primer)

[0078] The specific primer designed according to the mutant kdr L1014S type mutant gene (SEQ ID NO: ...

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Abstract

The invention discloses a kit for detecting the drug resistance of culex pipiens and a special primer thereof. The invention provides a primer for detecting kdr (knock-down resistance) gene mutation sites in culex pipiens, which is composed of a primer group A, a primer group B and a primer group C, wherein the primer group A is composed of a primer 1, a primer 2 and a primer 3; the primer group B is composed of the primer 1, the primer 2 and a primer 4; and the primer group C is composed of the primer 1, the primer 2 and a primer 5. In the invention, the experiment proves that by the quick, simple, convenient and sensitive method for detecting kdr mutation in culex pipiens, the site mutation of the detected sample can be accurately, effectively and directly detected, thus resistant individuals and sensitive individuals can be distinguished, and the genotypes of the individuals can be identified.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a drug resistance detection kit for Culex amphetaceus and special primers thereof. Background technique [0002] Culex pipiens complex belongs to the subgenus Culex pipiens of the genus Culex, which has a global distribution. It is generally believed that the composite group includes Cx.p.pipiens, Cx.p.quinquefasciatus, Cx.p.pallens and Cx.p.molestus ). The above four subspecies are distributed in my country. The types of this compound group are not only widely distributed, but also have great medical importance. It is one of the main targets of disease prevention and mosquito eradication in my country. Chemical control is still an important means of mosquito control at present and in the foreseeable future. However, the development of resistance seriously affects vector mosquito control. The resistance to pyrethroid insecticides of various mosquitoes in the Culex acutus comple...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 赵彤言汪中明李春晓董言德郭晓霞张映梅刘美德邢丹
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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