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Method and kit for rapidly detecting drug resistance of housefly to dimethyl dichloroviny phosphate (DDVP)

A drug-resistant, housefly technology, applied in the direction of material analysis by observing the impact on chemical indicators, and analysis by chemical reaction of materials, etc. take a long time, etc.

Inactive Publication Date: 2012-04-04
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, there are many methods such as biological assay methods, biochemical methods, immunological methods, molecular biological methods, etc. for the detection of the resistance of houseflies. Bioassay methods such as drip method and drug film method have great limitations
At present, no matter which method of determination is used, it takes a long time, and it is basically impossible to perform rapid detection on the spot, and the guidance for the use of pesticides is not timely enough.

Method used

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  • Method and kit for rapidly detecting drug resistance of housefly to dimethyl dichloroviny phosphate (DDVP)
  • Method and kit for rapidly detecting drug resistance of housefly to dimethyl dichloroviny phosphate (DDVP)
  • Method and kit for rapidly detecting drug resistance of housefly to dimethyl dichloroviny phosphate (DDVP)

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Experimental program
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preparation example Construction

[0030] 1 Preparation of Musca domestica crude enzyme source

[0031] Put 80 housefly adults 2-3 days after eclosion in a -80°C ultra-low temperature refrigerator for 30 minutes, take their heads with tweezers and put them into a 1.5mL centrifuge tube, add 1mL of ice-cold phosphate buffer (PB, 25mmol / L, pH=7.4, containing 0.1% Triton-100), homogenized, and the whole operation was performed on ice. The homogenate was centrifuged at 10,000×g at 4°C for 24 min, and the supernatant was taken as the enzyme source, and stored at 4°C for future use.

[0032] 2 Protein Quantification of Musca domestica Crude Enzyme Source

[0033] 2.1 Preparation of standard curve

[0034] Take 7 centrifuge tubes and operate in parallel according to Table 1.

[0035] Table 1 standard curve with protein solution preparation method

[0036]

[0037] After mixing evenly, add to the microtiter plate respectively, take No. 1 as the blank control, and measure the OD within 20min 595 Absorbance at nm,...

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Abstract

The invention discloses a method and a kit for rapidly detecting the drug resistance of a housefly to dimethyl dichloroviny phosphate (DDVP). Acetylcholinesterase (AChE) in the housefly is taken as a target of the DDVP, and can decompose a substrate acetylthiocholine (ATCh) iodide into an acetic acid and thiocholine iodide. The method specifically comprises the following steps of: performing color reaction of the thiocholine iodide and a sulphydryl color development agent 5,5-dithiobis(2-nitrobenzoic acid) (DTNB) to generate a yellow complex such as 5-mercapto-2-nitrobenzoic acid; when the AChE is not suppressed, namely the housefly is resistant, displaying yellow in the color reaction; and when the AChE is suppressed, namely the housefly is susceptible, displaying achromatic color in the color reaction. By the method and the kit, DDVP suppression concentration for distinguishing a susceptible strain from a resistant strain is made out by combining the activity, detected after the DDVP acts on the AChE, of the AChE in the housefly and a drug resistance level determined by housefly bioassay, and then the method for rapidly detecting the drug resistance of the housefly to the DDVP by utilizing a biochemiluminescence method is provided.

Description

technical field [0001] The invention relates to a rapid detection method and kit for the resistance of houseflies to organophosphorus insecticide DDVP, which is to detect the resistance of houseflies by means of biochemistry. Background technique [0002] At present, organophosphorus insecticides such as dichlorvos DDVP, fenitrothion, phoxim, and various compounding agents are widely used in the prevention and control of forestry and agricultural pests; space spraying for pest control in urban green spaces and sanitation pest control Organophosphorus pesticides are also used more. Under the long-term selection pressure of organophosphorus insecticides such as DDVP, houseflies in most areas have developed resistance to them. Therefore, although the dosage of the insecticides is gradually increased, the control effect is generally reduced and the environmental pollution is serious. Understanding the insecticide resistance of houseflies in a certain area can regulate the use o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 曹晓梅杨振洲孙宏迪张晓龙
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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