Preparation method and applications of cDNA chip for detecting drug resistance level of bactrocera dorsalis resistant to trichlorphon and beta-cypermethrin

A technology of high-efficiency cypermethrin and fruit fly, applied in biochemical equipment and methods, microbial determination/inspection, chemical library and other directions, can solve problems such as lack of reports, achieve simple and effective process, accurate real-time monitoring, delay drug resistance Effect

Active Publication Date: 2015-01-21
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still few reports on how many genes are involved in drug resistance, whether different drug resistance levels involve the same drug resistance genes, and how drug resistance genes participate in the process of drug resistance.

Method used

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  • Preparation method and applications of cDNA chip for detecting drug resistance level of bactrocera dorsalis resistant to trichlorphon and beta-cypermethrin
  • Preparation method and applications of cDNA chip for detecting drug resistance level of bactrocera dorsalis resistant to trichlorphon and beta-cypermethrin
  • Preparation method and applications of cDNA chip for detecting drug resistance level of bactrocera dorsalis resistant to trichlorphon and beta-cypermethrin

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Example 1 Construction of a cDNA chip for detecting resistance levels of Bactrocera dorsalis against trichlorfon and beta-cypermethrin

[0035] 1. Construction of SSH library of Bacteralis dorsalis resistant strains resistant to trichlorfon and beta-cypermethrin

[0036] (1) Breeding of resistant strains of Bactrocera dorsalis:

[0037] Use the sensitive strains kept in the laboratory as the parental F strains screened for drug-resistant strains 0 , reared under the same conditions indoors, and screened generation by generation with trichlorfon and beta-cypermethrin by using the drug film method.

[0038] Sensitive strain F 0 Selection criteria: the lethal middle concentration LC of trichlorfon to sensitive strains of Bactrocera dorsalis adults 50 =2.65mg / L; the lethal middle concentration LC of beta-cypermethrin to sensitive strains of Bactrocera dorsalis 50 =3.36mg / L.

[0039] Acquisition of resistant strains: After some generations of screening, trichlorfon wa...

Embodiment 2

[0054] Example 2 Detection of the drug resistance of Bactrocera dorsalis resistant strains by the cDNA chip prepared by the present invention

[0055] 1. Breeding of resistant strains of Bactrocera dorsalis

[0056] The sensitive strains kept in the laboratory were used as the parental F0 for the screening of drug-resistant strains. They were raised under the same conditions in the room, and were screened generation by generation with trichlorfon and beta-cypermethrin using the drug film method. The specific steps are the same as above. Results 3.21 times, 18.23 times and 69.5 times resistance to trichlorfon and 6.8 times, 16.2 times and 160.74 times resistance to beta-cypermethrin were obtained respectively. At the same time, the cross-resistance to trichlorfon of 160.74-fold strains resistant to beta-cypermethrin was determined to be 21.20 times.

[0057] 2. Extraction of total RNA from B. dorsalis resistant strains and sensitive strain samples

[0058] Total RNA was ex...

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Abstract

The invention discloses a cDNA chip for detecting the resistance level of Bactrocera dorsalis against trichlorfon and beta-cypermethrin, a production method and application thereof. The present invention utilizes the forward suppressive subtractive hybridization (suppression subtractive hybridization, SSH) method to respectively obtain different resistant strains of Bacteralis dorsalis drug resistance-specific up-regulated expression gene fragments, and then PCR amplifies each drug resistance-related specific expression gene, purifies and recovers 118 specific gene fragments related to resistance to trichlorfon of Bactrocera dorsalis and 103 specific gene fragments related to resistance to beta-cypermethrin of Bactrocera dorsalis were obtained. Using the above-mentioned specific resistance target gene fragments as DNA probes, a cDNA chip for the detection of Bacteralis dorsalis resistance to trichlorfon and beta-cypermethrin was constructed. The cDNA chip of the invention can simply and effectively realize the real-time monitoring of the drug resistance level of Bactrocera dorsalis.

Description

technical field [0001] The invention relates to the technical field of insect drug resistance detection. More specifically, it relates to a cDNA chip for detecting the resistance level of Bactrocera dorsalis against trichlorfon and beta-cypermethrin, its preparation method and application. Background technique [0002] Insect resistance was defined by the World Health Organization in 1957 as: "The phenomenon in which insects have the ability to tolerate and develop in their populations a dose that kills the majority of individuals in a normal population". Insect resistance to insecticides is an evolutionary phenomenon, an external manifestation of the continuous change of the internal genetic structure of insect populations under the action of insecticide selection, and a characteristic of insect populations, mainly including natural drug resistance, interaction drug resistance, negative cross-resistance, and multi-drug resistance. The consequences of insect resistance ar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C40B40/08C40B50/06
CPCC12Q1/6888C12Q2600/158
Inventor 曾玲姜建军陆永跃何晓芳梁广文温硕洋
Owner SOUTH CHINA AGRI UNIV
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