African swine fever virus triple fluorescent PCR detection kit and application thereof

Abstract

The invention discloses an African swine fever virus triple fluorescent PCR detection kit as well as a preparation method and application thereof. The detection kit comprises a fluorescent PCR reaction solution, positive control, negative control, an exogenous gene standard solution and a negative extraction control solution, wherein the fluorescent PCR reaction solution is prepared from upstreamand downstream primers and probes of a VP72 gene, an exogenous gene and an endogenous gene. According to the detection kit, the endogenous gene serves as internal quality control in the whole detection process, and is used for monitoring the whole process of sample collection, extraction and amplification; and the exogenous gene serves as external positive quality control in a nucleic acid extraction process, is added into a sample nucleic acid extraction process to be subjected to nucleic acid extraction along with the sample, and the nucleic acid extraction effect of the nucleic acid extraction kit is monitored according to the change of a PCR reaction Ct value of the exogenous gene, so that the accuracy of a detection result is improved, and false negative outcome is reduced.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis of animal pathogens, and relates to an African swine fever virus triple fluorescent PCR detection kit and a preparation method and application thereof. Background technique [0002] African swine fever (ASF) is an acute, febrile and highly contagious disease of pigs caused by African swine fever virus (ASFV). The disease has a short course of disease, high mortality, and is very harmful to the pig industry. It is required by the World Organization for Animal Health (OIE) to notify animal diseases, and it is also a type of animal disease that my country focuses on prevention and control. In 2018, my country reported the first ASF epidemic in Liaoning Province, and then it spread to other parts of the country, causing great economic losses to my country's pig industry. Since the clinical symptoms and pathological changes of ASF are difficult to distinguish from other diseases such as sw...

AI Technical Summary

Problems solved by technology

Since the clinical symptoms and pathological changes of ASF are difficult to distinguish from other diseases such as swine fever, and no effective vaccine is currently available, rapid and accurate laboratory diagnosis is of great significance

Classical detection and diagnosis methods such as blood adsorption test and virus isolation and identification have high requirements on the conditions of biosafety facilities, and are also limited in terms of test specificity and detection time.

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