Detection method for A, B, J subgroup avian leucosis viruses in avian infectious laryngotracheitis live vaccine

Abstract

The invention discloses a detection method for A, B, J subgroup avian leucosis viruses in an avian infectious laryngotracheitis live vaccine. The detection method comprises the following steps: diluting a commercial avian infectious laryngotracheitis live vaccine by adopting DMEM, then inoculating DF-1 cells, acting for 1 h at the temperature of 37 DEG C, then changing into DMEM containing 1%FBS, and culturing for 7 days; collecting cell culture supernatant, and carrying out avian leucosis virus p27 group specific antigen ALV-p27 detection; collecting cell cultures for an ALV-p27 positive sample and extracting cDNA, identifying A, B, J subgroup avian leucosis viruses by adopting a subgroup specific primer, and carrying out IFA detection on corresponding positive porocytes of the A, B, J subgroup avian leucosis viruses by adopting a subgroup specific monoclonal antibody. The detection method disclosed by the invention can enable live viruses to be proliferated in cells, makes up the defects of direct PCR detection and also has the characteristics of effectiveness, accuracy, reliability and the like and great significance on prevention and control of avian leucosis.

Description

Technical field [0001] The invention belongs to the field of avian leukemia virus detection, and particularly relates to a method for detecting avian leukemia virus of subgroups A, B, and J in a live vaccine of infectious laryngotracheitis. Background technique [0002] Avian leukosis virus (ALV) belongs to the retroviral family and belongs to the genus of retroviruses, which can cause benign, malignant and transmissible tumor diseases in poultry. ALV is divided into 11 subgroups of AK, of which 7 subgroups A, B, C, D, E, J, and K can naturally infect chickens. Subgroups A, B, and J are easily separated in clinical practice. Subgroup K It is a new subgroup discovered recently. Clinical infections and subclinical symptoms caused by ALV have caused huge economic losses to the world's poultry industry. In chicken flocks, the mortality rate caused by ALV infection is 1%-2%, and occasionally can reach 20% or even higher. In addition, ALV can also cause subclinical infections, reduc...

AI Technical Summary

Problems solved by technology

[0004] The detection method of A, B, J subgroup avian leukosis virus in commercially available commercial live vaccine mainly adopts the method of direct PCR, but the situation that the detection of direct PCR may be missed due to the low virus content; The target fragment amplified by PCR is a gene fragment rather than a complete live virus

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