A hrm non-labeled probe method for rapidly distinguishing prrsv classic strains and variant strains and its primers and probes

Abstract

The invention discloses an HRM (high-resolution melting) label-free probe method, a primer and a probe for quickly differentiating PRRSV (porcine reproductive and respiratory syndrome virus) classical strains and mutant strains. The HRM label-free probe method, the primer and the probe have the advantages that the primer and the probe are easy to operate; only the UN-Probe and fluorescent saturable dye need to be added into the strains before PCR (polymerase chain reaction) is carried out; the HRM label-free probe method, the primer and the probe are high in detection speed and throughput; complete operation procedures only take 3 hours, and accordingly required typing time can be greatly shortened; the HRM label-free method, the primer and the probe are low in cost, specific fluorescent label probes can be omitted, and the saturable dye cost of each sample is 1.6RMB; the HRM label-free probe method, the primer and the probe are high in accuracy and good in specificity and repeatability, the PRRSV classical strains and mutant strains can be accurately and quickly analyzed under a high-throughput condition, and accordingly the HRM label-free probe method, the primer and the probe are favorable to popularization and application in clinical practice.

Description

technical field [0001] The invention relates to a method for distinguishing classic virus strains and variant strains, in particular to an HRM non-labeled probe method for quickly distinguishing classic strains and variant strains of PRRSV, as well as primers and probes thereof. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is caused by porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV) infection. Reproductive disorders and respiratory diseases in piglets and finishing pigs. According to the difference of viral genome, PRRSV can be divided into two genotypes, namely European type and North American type. Although the prevailing strains of PRRSV include American type and European type, so far, the genotypes of PRRSV prevalent in my country are all North American types, including the classic strain (C-PRRSV) represented by the North American type PRRSV CH-1a strain. ) and mutant...

AI Technical Summary

Problems solved by technology

Although the above methods can distinguish between classic and mutant strains of PRRSV, traditional PCR detection techniques such as the One Step RT-PCR method require gel electrophoresis analysis of RT-PCR products, which can easily cause contamination of PCR products, resulting in false positives and poor sensitivity. Low

Fluorescence quantitative PCR has high sensitivity, but it is expensive, and two or more probes need to be synthesized at the same time

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