A method for detecting porcine viral diseases based on tem-pcr and gene chip

Abstract

The invention discloses a method for detecting pig viral diseases on the basis of TEM-PCR and a gene chip. The method comprises the steps that according to a target sequence, enriching multiplex-PCR, adopting four pairs of specific nested PCR primers and a probe, and adding a tag sequence which can be recognized by a pair of universal primers respectively on the 5 prime end of each inner primer. According to the method, a lot of defects of a traditional PCR method are overcome, the compatibility, specificity, flexibility and the like are all better than the traditional PCR method; the reaction conditions of Tem-PCR are optimized, and an efficient and accurate, and convenient and quick detection method is established; the background noise of the reaction is reduced, and the fund is greatly saved; an advanced Tem-PCR multiple amplification technology and a gene chip detection technology are combined for the first time, a system through which four etiologies can be detected on one piece of chip at the same time is established, and a foundation is laid for later molecular biology detection of important pathogenic microorganism.

Description

technical field [0001] The invention belongs to the technical field of swine fever detection, in particular to a method for detecting swine viral diseases based on TEM-PCR and a gene chip. Background technique [0002] African swine fever virus (ASFV), swine vesicular disease virus (SVDV), foot-and-mouth disease virus (FMDV), and porcine pseudorabies virus (PRV) are four important pathogens of swine diseases. Although there is no report of African swine fever in my country at present, due to the repeated reports of the occurrence of the disease and its possible serious consequences in neighboring Russia, it is necessary to establish a detection method early on to prevent the disease, and it can provide Future research on the disease provides a certain experimental basis. Porcine vesicular disease and foot-and-mouth disease have very similar clinical manifestations, and it is difficult to distinguish them from clinical symptoms. Therefore, establishing a rapid and accurate me...

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Problems solved by technology

Although there is no report of African swine fever in my country at present, due to the repeated reports of the occurrence of the disease and its possible serious consequences in neighboring Russia, it is necessary to establish a detection method early on to prevent the disease, and it can provide Provide a certain experimental basis for future research on the disease

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