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Method for detecting pig viral diseases on basis of TEM-PCR and gene chip

A viral disease and gene chip technology, applied in the field of swine viral disease detection based on TEM-PCR and gene chip, to achieve the effects of good flexibility, saving experimental costs, and reducing background noise

Active Publication Date: 2016-05-04
SICHUAN AGRI UNIV
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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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  • Method for detecting pig viral diseases on basis of TEM-PCR and gene chip
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  • Method for detecting pig viral diseases on basis of TEM-PCR and gene chip

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Embodiment Construction

[0043] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0044] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0045] Such as figure 1 Shown, the method for detecting porcine viral disease based on TEM-PCR and gene chip of the embodiment of the present invention may further comprise the steps:

[0046] S101: Preparation of DNA microarray: fix oligonucleotides on the substrate by in situ synthesis, use glass slides as the support of the gene chip, and before the polymerization reaction, the surface should be derivatized with hydroxyl or amino groups;

[0047] S102: Extract nucleic acid from ce...

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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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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6837C12Q1/701C12Q2537/143C12Q2563/107C12Q2565/501
Inventor 王印彭善珍杨泽晓姚学萍邬旭龙张鹏飞冷伊依肖璐任梅渗曾相杰罗忠永胡凌林星宇张博刘亚东蒙正群李桂黎王波
Owner SICHUAN AGRI UNIV
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