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Detection method for poison of CMV (Cytomegalovirus) and PVY (Potato Virus Y) of single-head tobacco aphids

A detection method and a poisonous technology, which is applied in the detection of PVY poisoning and single-headed aphid CMV, can solve the problems of high detection cost of PCR diagnostic method, difficulty in large-scale application and promotion, and ineffective detection, etc., to achieve detection cost Low, high sensitivity, easy operation effect

Inactive Publication Date: 2014-03-19
GUIZHOU UNIV
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Benefits of technology

The technology described by this patented method allows smoking cigarettes to be tested quickly without being detected harmful substances like nicotine. This makes it possible to use less expensive equipment than previous methods while still giving accurate results even at larger scales.

Problems solved by technology

This patented technical problem addressed in this patents relates to detecting various types of insect attacks through molecular techniques like polymerase chain reaction or DNA hybridization. However current methods require expensive equipment and can cause significant loss if they don't work well during commercial use over long periods.

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  • Detection method for poison of CMV (Cytomegalovirus) and PVY (Potato Virus Y) of single-head tobacco aphids
  • Detection method for poison of CMV (Cytomegalovirus) and PVY (Potato Virus Y) of single-head tobacco aphids

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Embodiment Construction

[0023] Embodiments of the present invention: detection method for single-headed tobacco aphid CMV, PVY poisoning,

[0024] 1) Put the single-headed worm into a 0.2mL PCR tube with 50μl carbonate buffer, mash it with a toothpick, and centrifuge at 8000r / min for 3min;

[0025] 2) Take 3 μl of the supernatant and apply it to the nitrocellulose membrane, dry it at room temperature, and repeat the sample application 3 times.

[0026] 3) Immerse the spotted nitrocellulose membrane in 5% skimmed milk powder blocking buffer, and block at 37°C for 60 minutes;

[0027] 4) Wash the blocked nitrocellulose membrane 3 times with washing solution, 3 minutes each time; add the polyantibody solution of the virus to be detected, mix the antibody and PBS buffer at a volume ratio of 1:200, and incubate at 37°C for 1.5 h;

[0028] 5) Wash the antibody-reacted nitrocellulose membrane with washing solution for 3 times, each time for 3 minutes; immerse the nitrocellulose membrane in the horseradish...

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Abstract

The invention discloses a detection method for poison of CMV (Cytomegalovirus) and PVY (Potato Virus Y) of single-head tobacco aphids. The detection method mainly adopts reagents and materials including CMV and PVY primary antibodies, a corresponding enzyme label second antibody, a sample diluting solution, a washing solution and a developing solution. A detection plate is a nitrocellulose membrane (NCM) coated with CMV and PVY virus protein antigens carried by the tobacco aphids; an enzyme conjugate working solution is a goat anti-rabbit antibody coated with the CMV and PVY primary antibodies (goat anti-rabbit) and horse radish peroxidase. According to the detection method, the single-head tobacco aphids are subjected to a series of treatment and an antigen and antibody reaction basic principle is utilized to detect that whether the tobacco aphids have poison or not in time. The detection method has the active effects of strong specificity, high sensitivity, simplicity in operation and low detection cost; the detection method is suitable for being popularized and applied in fields in a large scale and has a wide application prospect.

Description

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Claims

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Application Information

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Owner GUIZHOU UNIV
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