Real-time fluorescence quantification PCR detection method for avian infectious laryngotracheitis virus

Abstract

The invention discloses a real-time fluorescence quantification PCR (real-time PCR) detection method for an avian infectious laryngotracheitis virus, relating to the molecular biology field. The real-time fluorescence quantification PCR detection method aims to the gB gene of the avian infectious laryngotracheitis virus to design and synthesize a pair of primers, namely PF: 5'-CAATGGCTTCGGAGAAAGAG-3' and PR: 5'-GGCAATCCTGATCCCATCTA-3'. By PCR amplification and standard product preparation, a fluorescence quantification PCR method for detecting the avian infectious laryngotracheitis virus is constructed; the specificity is stronger; the sensitivity is high; the operation is simple and fast; the real-time detection is performed on the PCR progress; the qualitative and quantitative analysis can be performed on each cycle; and the constructed fluorescence quantification PCR method for the avian infectious laryngotracheitis virus can be used for detecting the virus content in tissue.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a detection method for chicken infectious laryngotracheitis virus. Background technique [0002] Infectious laryngotracheitis virus (ILTV) is a member of the herpesviridae family. The disease mainly affects poultry and can cause chicken infectious laryngotracheitis (AILT) is an acute, contact upper respiratory infectious disease. It is characterized by dyspnea, coughing, and expectoration of an exudate containing blood samples. During autopsy, swelling, hemorrhage and erosion of the larynx and tracheal mucosa can be seen. The case fatality rate is above 20%. After the disease was first reported in the United States in 1925, it has now spread to many chicken farming areas in the world. The disease spreads quickly and has a high mortality rate. It occurs and prevails in many areas in my country, endangering the development of the chicken industry. At present, there...

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Problems solved by technology

[0004] The technical problem to be solved in the present invention is to provide a real-time fluorescence quantitative PCR detection method for chicken infectious laryngotracheitis virus in view of the deficiencies of the above-mentioned prior art, which solves the problem that traditional PCR methods are not specific, prone to false positives, and cumbersome to operate , easy to produce PCR pollution, can only semi-quantitative or qualitative analysis of the final product of the reaction, with strong specificity, high sensitivity, simple and fast operation, less prone to false positives, less likely to produce PCR pollution, and real-time detection of the PCR process , perform quantitative and qualitative analysis on each cycle, and the analysis results can be directly read out by the computer

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