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Dual SYBR Green I real-time fluorescence PCR detection primer and method for porcine pseudorabies virus and porcine circovirus type 2

A technology of porcine pseudorabies virus and porcine circovirus, applied in the direction of microorganism-based methods, biochemical equipment and methods, fluorescence/phosphorescence, etc., can solve the problems of subclinical infection and no unified diagnostic method, and achieve good sensitivity , Facilitate the effect of identification and diagnosis

Inactive Publication Date: 2012-08-29
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

As a result, so far, there is no unified and rapid diagnostic method, preventive vaccines and specific drugs for the disease, and the disease is sometimes subclinical infection, so it is very important to make a good diagnosis of the disease

Method used

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  • Dual SYBR Green I real-time fluorescence PCR detection primer and method for porcine pseudorabies virus and porcine circovirus type 2
  • Dual SYBR Green I real-time fluorescence PCR detection primer and method for porcine pseudorabies virus and porcine circovirus type 2
  • Dual SYBR Green I real-time fluorescence PCR detection primer and method for porcine pseudorabies virus and porcine circovirus type 2

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Embodiment

[0030] The sequences of the primers for detection of porcine pseudorabies virus and porcine circovirus type 2 double SYBR Green I real-time fluorescent PCR are as follows:

[0031] The sequence of the porcine circovirus type 2 primer is as follows:

[0032] Upstream primer P1: 5′-TAA CTA CTC CTC CCG CCA TAC-3′;

[0033] Downstream primer P2: 5'-GCC TAC GTG GTC TAC ATT TCC-3'.

[0034] The sequence of the porcine pseudorabies virus primer is as follows:

[0035] Upstream primer P3: 5'-CAACCC GCTCGTGCTC-3';

[0036] Downstream primer P4: 5'-GCT GCT CCT CCA TGT CCT-3'.

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Abstract

The invention discloses a dual SYBR Green I real-time fluorescence polymerase chain reaction (PCR) detection primer and a dual SYBR Green I real-time fluorescence PCR detection method for porcine pseudorabies virus and porcine circovirus type 2. The primer is designed and synthesized, and the dual SYBR Green I real-time fluorescence PCR detection method for the porcine pseudorabies virus and the porcine circovirus type 2 by using the primer comprises the following steps of: extracting deoxyribose nucleic acid (DNA) of a sample; and detecting the sample by using an SYBR Green I real-time fluorescence PCR reaction system and an SYBR Green I real-time fluorescence PCR amplification program. By the primer and the method, two viruses, namely the porcine pseudorabies virus and the porcine circovirus type 2 can be detected simultaneously, and porcine parvovirus, classical swine fever virus, porcine reproductive and respiratory syndrome virus and swine influenza virus cannot be detected. The primer and the method have the characteristics of higher sensitivity, repeatability and stability, and contribute to the identification and the diagnosis of pregnant sow reproductive disturbance virusdisease.

Description

technical field [0001] The invention relates to a real-time fluorescent PCR detection method, in particular to primers and a method for double SYBR Green I real-time fluorescent PCR detection of porcine pseudorabies virus and porcine circovirus type 2. Background technique [0002] Infection source and natural reservoir host of porcine pseudorabies virus (PRV). Pigs infected with PRV often cause disease in newborn piglets, and the mortality rate can reach 100%. Porcine pseudorabies can present a typical latent infection. Pigs of any age can form a latent infection after being resistant to acute infection. The virus stays latent in the pig for life. Under certain conditions, the latent virus can be activated, causing recurrent infection and spreading the virus outward. This mechanism makes it difficult to eradicate pseudorabies. Porcine circovirus type 2 (PCV2) is pathogenic and mainly causes a newly discovered infectious disease - multisystemic wasting syndrome in weaned pi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64C12R1/93
Inventor 魏战勇胡慧陈红英崔保安李明凤郭显坡贾艳艳
Owner HENAN AGRICULTURAL UNIVERSITY
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