Primer and probe of west nile virus and real time RT-PCR detection reagent kit with one-step method

Abstract

The invention relates to a group of primers and probes and a corresponding kit, including a pair of primers which can simultaneously detect West Nile virus and the reference gene and two TaqMan probes against the West Nile virus and the reference gene, so as to reduce the number of the primers and the probes in a PCR reaction system from the usual 6 to 4, further to realize the high efficient amplification of the West Nile virus and the reference gene in the same tube, thus realizing the detection of the target genes of the different species, having strong universality, reducing the number of the primers and the probes, the cost of the reagents and the operation steps, and reducing the pollution chances which need to be avoided generally in a PCR experiment. The kit of the invention is applicable to the detection of the West Nile virus in the scientific research or the clinical aspect, especially the monitoring of the West Nile virus in the prevention and the control works of viral diseases.

Description

technical field [0001] The patent of the invention belongs to the field of biotechnology, and relates to the detection and quantitative analysis of West Nile virus RNA using RT-PCR method in scientific research, clinical and virus epidemiological monitoring and other work. Specifically, it is a primer and probe for West Nile virus and a one-step real-time RT-PCR detection kit. Background technique [0002] West Nile virus and human disease [0003] West Nile Virus (WNV) disease is a severe zoonotic disease, which was first discovered in the Nile region of northern Uganda in 1937 and got its name. In recent years, West Nile virus infection has occurred frequently in regions and countries such as Europe, the Middle East, West Asia, and Russia, and has become the most infectious disease affecting the United States in recent years. West Nile virus is a flavivirus transmitted by ornithophilic mosquitoes such as Culex mosquitoes, with birds as the main animal host. Humans, hors...

AI Technical Summary

Problems solved by technology

There are several problems in this way: 1. Since there are differences in the housekeeping gene sequences of different species, when detecting the target gene of different species, it is necessary to design primers and probes for the housekeeping gene for each species, which lacks versatility and increases The complexity and cost of the experimental operation; 2. When the target gene and the housekeeping gene are divided into two tubes for parallel detection, the cost of reagents and manpower is increased, and the chance of cross-contamination is also increased

If detected in the same tube, the PCR reaction efficiency will be reduced due to the interaction between the two sets of primers and probes, and the design difficulty of primers and probes will be increased

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