Primer and probe for Hendra virus as well as one-step real-time RT-PCR detection reagent case

Abstract

The invention relates to a group of primer and probe and a reagent kit thereof, which comprises a pair of primer which can simultaneously detect Hendra virus and reference gene, and two TaqMan probes aiming at Hendra virus and reference gene, and makes the number of primer and probe in a PCR reacting system be reduced to four from six, thereby realizing the amplification with high efficiency of Hendra virus and reference gene in the same tube, realizing the detection for target gene of various spices, having strong practicability, reducing the number of primer and probe and the cost of experimental reagent and operation steps, and reducing the contamination chances of PCR experiment which is usually needed to be avoided. The reagent kit of the invention is suitable to the detection of Hendra virus in scientific researches or in clinics, especially is suitable to the monitoring for Hendra virus in the viral diseases protection and control work.

Description

technical field [0001] The patent of the present invention belongs to the field of biotechnology, and relates to the detection and quantitative analysis of Hendra virus RNA using RT-PCR method in scientific research, clinical and virus epidemiological monitoring and other work. Specifically, primers and probes for Hendra virus and a one-step real-time RT-PCR detection kit. Background technique [0002] Hendra virus and human diseases [0003] Hendra virus (HeV) is a new paramyxovirus emerging in recent years, which can cause acute central nervous system and respiratory system lesions in humans and animals. Hendra virus, formerly known as equine measles virus (equinemobil-livirUs, EMV), was first discovered near Brisbane, Queensland, Australia in 1994, and occurred again in Mackay (Mackay) in 1995. A total of 15 racehorses and 2 people were killed on both occasions. Mainly presented with respiratory symptoms, but in Mackay's case neurological symptoms first appeared. Afte...

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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