Establishment of detecting method for four import-dairy-cow-carried viruses GeXP

Abstract

The invention aims at establishing a detecting method for multiple dairy-cow epidemic diseases based on a GeXP-multiple-genetic-expression genetic analysis system. The problems that when the multiple dairy-cow epidemic diseases are detected, the sensitivity of the serology method is low, only a single pathogene is detected through conventional PCR, and the cost of the gene chip method is high are solved. Four dairy-cow epidemic-disease RNA-positive-sample reference products are built; ten-fold continuous dilution reference products of RNA of the reference products serve as detected objects, and the sensitivity is detected to 10-100 copies; the dairy-cow epidemic disease is free of cross reaction with other dairy-cow epidemic diseases, and false negative is avoided; 600 parts of practical samples are detected, and the monitoring screening requirements of the multiple epidemic diseases of a large number of dairy cows are met.

Description

technical field [0001] The invention relates to the application field of biotechnology, in particular to the simultaneous detection and identification of four imported cow virus diseases. Background technique [0002] Foot and Mouth Disease (FMD), Bovine Viral Diarrhea (BVD), Bluetongue (BT) and Akabane Disease (AKA) are four 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 carr...

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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