Development and application of multiple fluorescence RT-PCR detection method for foot-and-mouth disease, vesicular stomatitis and swine vesicular disease

Abstract

Based on a highly conserved domain of a foot-and-mouth disease virus 3D protein coding gene, a vesicular stomatitis virus N protein coding gene and a swine vesicular disease virus VP1 protein coding gene, the invention designs a specific primer and a probe and develops a multiple fluorescence RT-PCR detection method used for simultaneously detecting the three animal viruses. The detection method can detect 102 copied plasmids containing target amplification sequences in 1 h. The method has high sensitivity and good specificity, and can achieve rapid high-throughput fluorescence RT-PCR detection for single-virus infection or mixed infection by a plurality of viruses.

Description

technical field [0001] The invention relates to the application field of biotechnology, especially for simultaneous detection and identification of foot-and-mouth disease, vesicular stomatitis and swine vesicular disease. Background technique [0002] In the past 10 years, under the promotion of the government and the pull of processing and consumption demand, China's dairy industry has made great progress, and gradually developed from a poor milk country to a big dairy country. First of all, the number of imported dairy cows is increasing year by year. From 1997 to 2007, the number of dairy cows in China increased by 4.84 times; in 2009, 45,000 dairy cows were imported, in 2010, 80,000 dairy cows were imported, and in 2011, 100,000 dairy cows were imported. Secondly, the level of large-scale breeding of domestic dairy cows has been greatly improved. In 2010, the stock of more than 100 dairy cows in the country accounted for 28.48% of the country's total, an increase of 8....

AI Technical Summary

Problems solved by technology

The 3 vesicular diseases are clinically indistinguishable and must be differentially diagnosed by laboratory testing

At present, pathogen isolation and identification and conventional serological methods are mostly used for the diagnosis of the three diseases, which takes a long time, and PCR technology can also be used for detection, but there is no stable and operable method that can simultaneously identify the three viruses method

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