Detection method of exogenous avian leukemia virus

Abstract

The invention discloses a detection method of exogenous avian leukemia virus. The method comprises the following steps: carrying out digestion and dilution on well-grown DF1 cells, carrying out subculture in a cell culture plate, and culturing at 37 DEG C in 5% CO2 for 25-35 minutes; when 45-55% of cells are deposited on the bottom of the cell culture plate, sequentially adding plasma of the detected chicken into every hole of the cell culture plate along the wall, culturing over night, discarding the raw liquor in the cell culture plate, adding maintenance media containing 1% of fetal calf serum, continuing culturing for 7-8 days, and cryopreserving the culture product; and defreezing the culture product, uniformly mixing by slightly blowing and beating, adding the culture product into a P27 antigen reaction plate, detecting the P27 antigen, and judging whether the detected chicken is infected by the exogenous avian leukemia virus. Compared with the traditional process, the method disclosed by the invention can detect the same batch samples at higher positive rate, and has the characteristics of higher detected S/P average value and obvious differences; and the method has equal stability and accuracy, but has higher sensitivity.

Description

technical field [0001] The invention belongs to the technical field of microorganism separation and detection, and more specifically, the invention relates to a detection method of exogenous avian leukosis virus. Background technique [0002] Avian leukemia (AL) is a general term for a variety of tumor diseases caused by avian leukemia virus (ALV) and mainly characterized by malignant proliferation of hematopoietic cells. ALV can infect all kinds of chicken flocks. There are vertical transmission, horizontal transmission, insects, needles, vaccine contamination and other transmission modes, among which vertical transmission is the main infection mode. At present, there is no effective vaccine to prevent and control the disease, and there is no effective drug treatment. The only way to control the disease from the source is to cut off the source of ALV infection in the breeder farm through purification measures. [0003] At present, the purification of avian leukemia is main...

AI Technical Summary

Problems solved by technology

[0004] However, in the traditional virus isolation method, the plasma is not inoculated until the DF1 cells grow to 70-80% monolayer. At this time, the cells are in good condition and very stable, which increases the difficulty for the virus to invade the cells.

The virus did not fully participate in the attachment, growth, and division of DF1 cells, and the medium was changed after only 2 hours of incubation after inoculation. Some viruses did not invade the cells and could not reproduce, resulting in a low positive rate of P27 antigen detection

In addition, the traditional virus isolation method is complicated to operate, requiring steps such as culturing monolayer cells and incubation, and the technology is difficult to master

Images