Method for quickly detecting duck viral enteritis by real-time fluorescence quantitative PCR and kit thereof

Abstract

The invention provides a method for quickly detecting duck viral enteritis by real-time fluorescence quantitative PCR. The method selects a gene fragment of duck viral enteritis UL31 to design a primer and a probe, and order-checking results on an amplified fragment proves that homologies of the amplified fragment and the gene fragment are both 100 percent without a cross reaction, thereby eliminating problems of virus variation and false negative caused by region factors, and improving detectable positive rate and compatibility of the method. The method constructs a quantitative positive quality control standard (recombinant plasmid pMD18-T-DEV76bp), establishes a standard curve of fluorescence quantitative detection, and can minimally detect 2.1*10 degrees copy / mu liter. The assembled standard kit reacts under a standard reaction condition, and determines according to accurate quality control standard and determination standard. The method has the advantages of strong specificity, high sensibility, safe quantitative positive quality control stand, simple operation and low cost, is applicable to large-scale quarantine inspection, port quarantine inspection, epidemiological investigation and laboratory quick diagnose of field duck viral enteritis.

Description

technical field [0001] The invention belongs to the category of molecular biology, specifically a method and a kit for rapidly detecting duck viral enteritis by using fluorescent PCR technology. Background technique [0002] In the past ten years, my country's duck industry has developed rapidly. According to the statistics of the United Nations Food and Agriculture Organization (FAO) in 2002, my country's duck population is about 661 million, accounting for 69.7% of the world's total. It can be seen that my country is the largest duck producer . Moreover, with an annual rate of 10% to 15%, newly-built large-scale farms are gradually increasing, and the annual output value of duck products is close to 30 billion yuan, and they are exported to the European Union, Southeast Asia, Japan, South Korea and other places. With the further development of the duck industry, the space for duck products will continue to expand, so the duck industry has huge potential. However, due to t...

AI Technical Summary

Problems solved by technology

However, due to the introduction of breeding ducks or eggs through various channels, frequent exchanges in the domestic poultry product market, lack of management experience in intensive breeding, improper epidemic prevention measures, and serious environmental pollution, various diseases of ducks continue to occur and become more and more complicated. The more serious hazards are still viral infectious diseases, which account for 15% to 20% of duck deaths every year, seriously restricting the sustainable and stable development of my country's duck industry

Images