Fluorescent probe primer group and kit for African swine fever virus P72 gene, and application thereof

Abstract

The invention relates to a fluorescent probe primer group and a kit for an African swine fever virus P72 gene, and application thereof. The primer comprises an upstream primer ASFV-F: 5 '-CCACGGGAGGAATACCAA-3', the downstream primer ASFV-R is 5 '-GCAGATGCCGATACCACA-3'; a fluorescent probe ASFV-P, containing a fragment 5 '-TCATTAACGTATCCAGAGCAAGA-3', the 5' end of the fluorescent probe ASFV-P is marked with a fluorescent reporter group, and the 3 ' end of the fluorescent probe ASFV-P is marked with a fluorescent quenching group. The kit comprises a fluorescent probe primer group, a positive control, a negative control and a PCR amplification solution. The application specifically comprises the following steps: 1) extracting total DNA of a sample to be detected; 2) taking the total DNA obtained in the step 1) as a template, preparing a fluorescence reaction system by using the kit, and carrying out fluorescence amplification; and (3) establishing a standard curve by utilizing positive control, analyzing a fluorescence amplification product, and judging whether the sample to be detected is positive or negative according to an amplification reaction result. Compared with the prior art, the primer group has the advantages that all current epidemic ASFV strains can be identified to the maximum extent, and the sensitivity of the primer is improved to 1.15 * 10 < 1 > copise / [mu]L.

Description

technical field [0001] The invention belongs to the field of animal pathogen detection, and in particular relates to a fluorescent probe primer set, kit and application of African swine fever virus P72 gene. Background technique [0002] African swine fever (ASF) is an acute, febrile, highly contagious disease of pigs caused by African swine fever virus (ASFV). Domestic pigs and wild boars are generally susceptible, and soft ticks are the storage host and medium of ASFV. The incubation period of ASFV is 5-15 days. Once an outbreak occurs, the course of disease is short and the case fatality rate is high, which can reach 100%. The clinical symptoms of African swine fever are complex and difficult to distinguish from other diseases, mostly manifested as high fever, skin congestion, abortion, edema and organ hemorrhage. There is no effective treatment after the onset of African swine fever, and the African swine fever virus has a complex immune evasion mechanism, lacks typica...

AI Technical Summary

Problems solved by technology

There is no effective treatment after the onset of African swine fever, and the African swine fever virus has a complex immune evasion mechanism, lacks typical neutralizing antibodies, and has no effective vaccine for epidemic prevention. Therefore, a rapid, sensitive and accurate detection method is established appears to be particularly important

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