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Multiplex PCR (polymerase chain reaction) detection of porcine circovirus type II, porcine parvovirus and porcine pseudorabies virus and special primers thereof

A technology of porcine pseudorabies virus and porcine circovirus, which is applied in the biological field, can solve problems such as time-consuming, laborious, uneconomical, and inconvenient, and achieve good application prospects, save detection time, and save detection costs.

Inactive Publication Date: 2012-01-25
山东省动物疫病预防与控制中心
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Commercial PCR kits for PCVII, PPV, and PRV are all single-item PCR, and one kit can only detect one virus in one test. For the same pig disease material mixed with PCVII, PPV, and PRV, it is necessary to confirm the pathogen. Three kinds of kits are required and three PCR experiments are required, which is not only time-consuming and labor-intensive, but also has a high detection cost, which is neither economical nor fast

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  • Multiplex PCR (polymerase chain reaction) detection of porcine circovirus type II, porcine parvovirus and porcine pseudorabies virus and special primers thereof
  • Multiplex PCR (polymerase chain reaction) detection of porcine circovirus type II, porcine parvovirus and porcine pseudorabies virus and special primers thereof
  • Multiplex PCR (polymerase chain reaction) detection of porcine circovirus type II, porcine parvovirus and porcine pseudorabies virus and special primers thereof

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Embodiment 1

[0053] Embodiment 1, circovirus II (PCVII), porcine parvovirus (PPV) and porcine pseudorabies virus (PRV) multiple PCR detection method construction

[0054] 1. Primer design

[0055] Part of the known sequences of PCVII (Genbank No. EF524528), PPV (Genbank No. AY502114), and PRV (Genbank No. AF257079) were found in GenBank. Homology analysis was carried out on each gene region of the virus, and the whole gene of PCVII, the NS1 gene of PPV, and the gB gene of PRV were respectively selected as the target sequence for amplification of each virus. DNAMAN was used to design primers for the above conserved regions, and DNAstar was used to analyze the primer dimers of the three designed viral primers to avoid the formation of stable primer dimers between the primers. Select the following 3 pairs of primers whose amplified sequences are PCVII (1265bp), PPV (748bp), and PRV (342bp), and analyze the homology or complementarity of the amplified sequences of each pair of primers. Avoid...

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Abstract

The invention discloses a multiplex PCR (polymerase chain reaction) detection reagent of porcine circovirus type II, porcine parvovirus and porcine pseudorabies virus and special primers thereof. The invention provides the special primers for detecting the virus, and the special primers comprise a primer pair A, a primer pair B and a primer pair C, wherein the primer pair A comprises a primer 1 and a primer 2, the primer pair B comprises a primer 3 and a primer 4, the primer pair C comprises a primer 5 and a primer 6, and the nucleotide sequences of the primer 1, the primer 2, the primer 3, the primer 4, the primer 5 and the primer 6 are respectively as shown in the sequence 1, the sequence 2, the sequence 3, the sequence 4, the sequence 5 and the sequence 6 in a sequence table. Experiments prove that the multiplex PCR detection method of the porcine circovirus type II, the porcine parvovirus and the porcine pseudorabies virus provided by the invention is simple, convenient, fast, economical and efficient and has great application prospects in clinical diagnosis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to multiplex PCR detection of porcine circovirus type II, porcine parvovirus and porcine pseudorabies virus and special primers thereof. Background technique [0002] The PCR detection method of porcine circovirus II (PCVII), porcine parvovirus (PPV) and porcine pseudorabies virus (PRV) is currently a common method for rapid detection of these three pathogens, and is widely used in laboratory pathogen diagnosis. Compared with the isolation method, the PCR method is faster and easier to diagnose, and the time for virus isolation is relatively long, which is not conducive to rapid clinical diagnosis. Therefore, the current laboratory diagnosis of viral diseases mainly uses the PCR / RT-PCR method. At present, some PCVII, PPV, and PRV PCR kits have been commercialized and put into clinical application. [0003] At present, the multi-virus mixed infection of swine disease is quite serious. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12N15/11C12Q1/68
Inventor 李云岗杜兰荣张栋孙圣福兰邹然
Owner 山东省动物疫病预防与控制中心
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