Primer combination, probe combination, application of primer combination or probe combination to detection of porcine viruses, detection reagent, kit and detection method

Abstract

The invention discloses a primer combination, a probe combination, an application of the primer combination or the probe combination to detection of porcine viruses, a detection reagent, a kit and a detection method. According to the primer combination, the probe combination, the application of the probe combination to detection of porcine viruses, the detection reagent, the kit and the detectionmethod, a five-porcine-virus joint detection method is established, five major porcine disease single tubes of ASFV, FMDV, CSFV, HP-PRRSV and PRV wild viruses are rapidly detected, high-throughput screening of the five viruses can be realized, the detection efficiency can be improved, and the reagent consumption can be reduced. Therefore, the cost is saved, the detection time is shortened and theworkload is reduced. The invention provides the detection reagent capable of realizing joint detection of the five porcine viruses, and the optimized primer combination and the optimized probe combination are adopted, so that the reagent and the kit adopting the reagent can specifically and sensitively detect the five porcine viruses, namely the ASFV, FMDV, CSFV, HP-PRRSV and PRV wild viruses; andfive porcine virus infections of different types of nucleic acids can be simply, quickly and accurately detected in parallel in a single reaction tube, and DNA viruses and RNA viruses can be simultaneously detected by single-tube PCR.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a primer combination, a probe combination and their application in detecting porcine viruses, a detection reagent, a kit and a detection method. Background technique [0002] African swine fever (ASF) is an acute, febrile, highly contagious zoonotic disease of pigs caused by African swine fever virus (ASFV), characterized clinically by high fever and reticuloendothelial bleeding Characterized by high mortality and high mortality, the case fatality rate of susceptible pigs is as high as 100%. The World Organization for Animal Health (OIE) has listed African swine fever as a legally notifiable animal disease, and my country has listed it as a first-class animal disease. In August 2018, my country's first case of African swine fever was diagnosed, and then African swine fever broke out in many places, which brought a heavy blow to my country's pig breeding industry. ...

AI Technical Summary

Problems solved by technology

Although some scholars have established five or more target fluorescent quantitative PCR methods, they are all based on dye-based fluorescent PCR. Due to the problem of false positives, it is difficult to apply to clinical sample detection.

[0010] At present, there are many fluorescent quantitative PCR methods that can detect ASFV, FMDV, CSFV, HP-PRRSV and PRV wild virus alone, but there are no reports on the TaqMan probe five-fold fluorescent quantitative PCR method that can detect the above five porcine viruses simultaneously.

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