Primers and assays for gexp multiplex rapid detection for Akabane virus, bovine viral diarrhea virus and bluetongue virus

Abstract

This study intends to establish a detection method for multiple dairy cow diseases based on the GeXP multiple gene expression genetic analysis system to solve the problems of low sensitivity of serological methods in the detection of various dairy cow diseases, conventional PCR can only detect a single pathogen, and the high cost of gene chip methods. Construct 3 kinds of dairy cow disease RNA positive sample reference products; 10-fold serial dilution of the reference sample RNA is used as the detection object, and the sensitivity is to detect 100 copies; there is no cross-reaction with other dairy cow diseases, and there is no false negative; 600 actual samples are tested , to meet the monitoring and screening requirements of a large number of dairy cows for various diseases.

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), Bovine Viral Diarrhea (BVD) 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 epidemiolo...

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

Images