PCR-HRM primer and method for rapidly distinguishing wild strain and vaccine strain of canine parvovirus

Abstract

The invention discloses a PCR-HRM primer and a method for rapidly distinguishing a wild strain and a vaccine strain of canine parvovirus. The method comprises the following steps of: extracting a virus DNA from a sample as a template, carrying out PCR amplification by utilizing designed two pairs of specific primers and fluorescent saturated dye, carrying out HRM analysis on the detected sample respectively with a wild strain standard sample and a vaccine strain standard sample as contrast, and determining the type of the canine parvovirus. With the adoption of the method, the operation is simple, and only is the fluorescent saturated dye added before the PCR; the detection speed is high, the flux is high, the whole operation process only needs 3 hours, and cell culture of viruses is not needed, and thus the time needed by distinguishing detection is greatly shortened; the cost is low, a specific probe is not needed, and the fluorescent saturated dye is cheap and easily obtained; as the accuracy is high, the specificity and the repeatability are good, and analysis can be accurately and rapidly carried out with high flux, the method is beneficial to popularizing and applying in clinical practices.

Description

technical field [0001] The invention relates to a method for identifying virus vaccine virus and wild strain, in particular to a PCR-HRM primer and method for quickly distinguishing canine parvovirus wild strain and vaccine strain. Background technique [0002] Canine parvovirus (CPV) is a single-stranded small DNA virus, which is the main cause of acute hemorrhagic gastroenteritis in dogs and acute myocarditis in puppies. In 1977, Eugster and Nairn first isolated canine parvovirus from the feces of dogs with hemorrhagic enteritis and named it CPV-2. Since the first isolation of CPV-2 by Eugster et al., the antigenicity of CPV has changed over time, and new CPV genotypes and antigenic types have emerged continuously, and have spread widely around the world. Currently known CPV genotypes include CPV-2a, CPV-2b, and CPV-2c. The original CPV-2 genotype has been replaced by a new antigen variant strain, and the original CPV-2 strain can only be treated with attenuated live vacc...

AI Technical Summary

Problems solved by technology

At present, the main method for distinguishing canine parvovirus vaccine strains from wild virus is fluorescent quantitative PCR (MGB probe). Although this method can accurately distinguish canine parvovirus vaccine strains from wild strains, it requires three or more pairs of probes at the same time. Expensive, limiting its practical application in production

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