GeXP rapid multi-detection primer for detecting akabane viruses, foot and mouth disease viruses and blue tongue viruses and detecting method

Abstract

The invention tries to establish a multi-cow-disease detecting method based on a GeXP multi-gene expression genetic analysis system. The problems that when various cow diseases are detected, a serological method is low in sensitivity, only a single pathogen can be detected through a conventional PCR, and a gene chip method is high in cost are solved. Three cow disease RNA positive sample reference products are established. A 10-fold continuous diluted reference product of the reference product RNA serves as a detecting object, and 100 copies can be detected in terms of sensitivity; no cross reaction or false negative can happen between the cow diseases and other cow diseases, 600 actual samples are detected, and the requirements for monitoring and screening a large number of cow diseases are met.

Description

technical field [0001] The invention relates to the application field of biotechnology, in particular to the simultaneous detection and identification of three imported cow virus diseases. Background technique [0002] Akabane Disease (AKA), Foot and Mouth Disease (FMD) and Bluetongue (Bluetongue, BT) are three viral diseases that require isolation and quarantine for imported dairy cows. In recent years, as the national demand for high-quality dairy products has increased, the number of imported dairy cows has increased year by year, which has brought a certain amount of work pressure and financial burden to animal disease testing laboratories at dairy cow entry ports. Therefore, it is of great significance to establish a high-throughput rapid detection method that can simultaneously differentially diagnose multiple dairy cow diseases to reduce the workload of the inspection laboratory at the port of entry of dairy cows, reduce the cost of inspection, and carry out epidemiol...

AI Technical Summary

Problems solved by technology

Virus isolation and culture is the gold standard for diagnosis, but there are problems such as long test period and cumbersome operation; virus neutralization test requires specific standard serum or virus strain, which has certain limitations in practical application; imported enzyme-linked immunosorbent immunoassay Kits are generally expensive; molecular biology methods such as ordinary PCR and fluorescent quantitative PCR are all aimed at the detection of a single virus, which cannot meet the detection needs of differential diagnosis of mixed infections with multiple pathogens

Multiplex PCR technology has been applied to the differential diagnosis of various pathogens, but due to the problem of amplification preference, it is impossible to realize high-throughput detection in the true sense

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