Method for detecting dengue virus in mosquito by immunofluorescence technique

Abstract

The invention provides a method for detecting dengue virus in mosquitoes by using an immunity fluorescent technology. DEN-2 virus from artificially infected mosquito is taken, detection is carried out respectively by cell culturing inoculating and tablet immunity fluorescent way at the head of the mosquito, and the body of the mosquito is detected to carry or not virus by tablet immunity fluorescent way at the head of the mosquito. In the invention, a single blood-fully infected aedes albopictus is frozen under -20 centigrade, the head of the aedes albopictus is cut down under a dissecting microscope by a surgical knife, and the head is placed on a slide and extruded by a dissecting needle, so that the substance in the head flows out and is evenly coated on the slide; the slide is aired dry and fixed by pre-cooled acetone for 10 min.; mouse anti DEN-2 virus IgG diluted by PBS is dripped on the slide; the slide is then placed in a wet box, incubated for 30 min under 37 centigrade; then the slide is washed 3 times by PBGS, aired dry; goat anti-mouse IgG-FITC that is diluted by Evans blue PBS to operation concentration is dripped on the slide; the slide is then incubated for 30 min under 37 centigrade, washed by PBS; the dyed result is observed under a fluorescent microscope. The invention allows earlier diagnosis or timely discovering of positive mosquito media, so as to control the spreading of dengue.

Description

Technical field: [0001] The invention relates to a method for virus detection, in particular to a method for detecting dengue virus in mosquitoes by immunofluorescence technology. Background technique: [0002] Dengue fever and dengue hemorrhagic fever (DF / DHF) are arbovirus diseases caused by four types of dengue viruses of the genus Flavivirus, which are widely prevalent in tropical and subtropical regions of the world, and are most serious in Southeast Asian countries. Larger epidemics have also occurred in Hainan and the coastal areas of Guangdong. At present, the method of detecting dengue virus infection is still inseparable from animal inoculation or mosquito cell culture inoculation and isolation. The experimental results show that the detection of dengue virus by cell inoculation, culture and isolation is a more reliable method for the diagnosis of the pathogen of epidemic bacteria, but it is time-consuming and cumbersome. , requires strict experimental conditions,...

AI Technical Summary

Problems solved by technology

At present, the method of detecting dengue virus infection is still inseparable from animal inoculation or mosquito cell culture inoculation and isolation. The experimental results show that the detection of dengue virus by cell inoculation, culture and isolation is a more reliable method for the diagnosis of the pathogen of epidemic bacteria, but it is time-consuming and cumbersome. , requires strict experimental conditions, and the results can be produced within at least 1 week, failing to achieve the purpose of early and rapid diagnosis

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