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A qualitative and quantitative detection method of Periplaneta melanogaster densovirus and its application

A technology of densovirus and Periplaneta melanogaster, applied in the field of insect virus biopesticide detection, can solve the problems of high mortality and environmental pollution, achieve high sensitivity, ensure product quality, and achieve rapid qualitative and quantitative detection effects

Active Publication Date: 2016-05-04
WUHAN UNIOASIS BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main method of preventing and controlling cockroaches for many years is to use chemical pesticides, which has caused the cockroaches to increase their drug resistance. There have been many reports on microbial control of cockroaches. Among them, Periplaneta black-breasted densovirus isolated from naturally diseased cockroaches has very good infectivity to cockroaches, with a mortality rate as high as 99%.

Method used

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  • A qualitative and quantitative detection method of Periplaneta melanogaster densovirus and its application

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Effect test

Embodiment 1

[0034] A qualitative and quantitative detection method for detecting Periplaneta melanogaster densovirus, the steps are as follows:

[0035] 1. Virion Purification

[0036] Densovirus samples of Periplaneta melanogaster (provided by Wuhan Wuda Oasis Biotechnology Co., Ltd.) were suspended in phosphate buffered solution (PBS), homogenized and centrifuged at 5000r / min for 3min. Add ammonium sulfate 350g / L to the supernatant (referring to the final concentration after adding ammonium sulfate), and the saturation of the ammonium sulfate aqueous solution is 55%. Centrifuge for 50 min, add PBS to the precipitate to resuspend, after homogenization, centrifuge at 15000r / min for 20min, centrifuge the supernatant at 40000r / min for 2h, take the precipitate. After the homogenate was centrifuged, the supernatant was subjected to a 40% sucrose single gradient, centrifuged at 40,000r / min for 5h, the pellet was suspended in PBS, homogenized, and centrifuged at 12,000r / min for 20min to obtain...

Embodiment 2

[0057] A kind of qualitative and quantitative detection method of Periplaneta melanogaster densovirus is applied in the detection of cockroach virus cockroach bait, and its application process is as follows:

[0058] The source of biological cockroach bait (provided by Wuhan Wuda Oasis Biotechnology Co., Ltd.), ≥6000 pcs / g.

[0059] Step is with embodiment 1

[0060] The content of biological cockroach bait is 7.1×10 4 pcs / g.

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Abstract

The invention relates to a qualitative and quantitative detection method for a Periplaneta fuliginosa densonucleosis virus and application. The method comprises the following steps: purification of a virion; identification of deposition characteristics of the virion; measurement of the morphology of the virion; extraction of viral nucleic acid; measurement of molecular weight of the nucleic acid through electrophoresis; and counting in virtue of an electron microscope (including sample treatment and counting). The detection method provided by the invention has the advantages of high detection accuracy, short detection time, good specificity and stability and suitability for qualitative and quantitative detection of the virus.

Description

technical field [0001] The present invention relates to the technical field of insect virus biopesticide detection, and more specifically to a qualitative and quantitative detection method and application for detecting Periplaneta melanogaster densovirus. The most important thing is that it can accurately detect the densovirus of Periplaneta melanogaster qualitatively and quantitatively. Background technique [0002] Cockroach scientific name cockroach, belongs to Insecta (Insecta), Orthoptera (Blattaria), Blattidae (Blattidae). Cockroach is a kind of very ancient insect, appeared as early as 300 million years ago, known as the title of living fossil. At the same time, cockroaches are one of the important sanitary pests that endanger human health. Its harm is no less than that of mosquitoes, flies and mice. Bacteria, Pseudomonas aeruginosa, Escherichia coli and other bacteria and ten kinds of parasite eggs such as hookworm, roundworm, whipworm, etc., can be stored in the b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/04G01N15/02G01N21/33G01N27/447
Inventor 陈娇尹宜农
Owner WUHAN UNIOASIS BIOLOGICAL TECH CO LTD
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