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Multiplex real-time fluorescence PCR (polymerase chain reaction) detection primer and method for porcine rabies virus, porcine parvovirus 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 poor sensitivity, inaccurate quantification, time-consuming and labor-intensive, etc., and achieve good sensitivity Sexuality, beneficial to the effect of differentiation and diagnosis

Inactive Publication Date: 2011-05-25
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0003] At present, there are many diagnostic methods for the above three viral diseases. These methods have the disadvantages of complicated operation, time-consuming and laborious, poor sensitivity and high negative rate; the PCR diagnostic technology that has been used in clinical practice has high sensitivity, strong specificity, rapid , simplicity, etc., but cannot be accurately quantified; based on TaqMan technology, the fluorescent quantitative PCR method can be quantified, but the cost is high, and specific probes need to be synthesized

Method used

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  • Multiplex real-time fluorescence PCR (polymerase chain reaction) detection primer and method for porcine rabies virus, porcine parvovirus and porcine circovirus type 2
  • Multiplex real-time fluorescence PCR (polymerase chain reaction) detection primer and method for porcine rabies virus, porcine parvovirus and porcine circovirus type 2
  • Multiplex real-time fluorescence PCR (polymerase chain reaction) detection primer and method for porcine rabies virus, porcine parvovirus and porcine circovirus type 2

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Embodiment

[0035] The sequences of the multiplex SYBR Green I real-time fluorescent PCR detection primers for porcine pseudorabies virus, porcine parvovirus and porcine circovirus type 2 are as follows:

[0036] The sequence of the porcine circovirus type 2 primer is as follows:

[0037] Upstream primer P1: 5′-TAA CTA CTC CTC CCG CCA TAC-3′;

[0038] Downstream primer P2: 5′-GCC TAC GTG GTC TAC ATT TCC-3′;

[0039] The sequences of porcine parvovirus primers are as follows:

[0040] Upstream primer P3: 5′-TGG GAG GGC TTG GTT AGA-3′;

[0041] Downstream primer P4: 5′-TGG TGG TGA GGT TGC TGA T-3′;

[0042] The sequence of the porcine pseudorabies virus primer is as follows:

[0043] Upstream primer P5: 5′-CGT GGA ACG AGC CCT TCA G-3′;

[0044] Downstream primer P6: 5'-AGA GCG GGT TGG CGA TGT-3'.

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Abstract

The invention discloses a multiplex SYBR Green I real-time fluorescence PCR (polymerase chain reaction) detection primer and method for porcine rabies virus, porcine parvovirus and porcine circovirus type 2. The primer is obtained through synthesis according to design. The multiplex SYBR Green I real-time fluorescence PCR detection method for detecting porcine rabies virus, porcine parvovirus and porcine circovirus type 2 by utilizing the primer comprises the following steps: extracting the DNA of a sample, and then, detecting the sample by utilizing a SYBR Green I real-time fluorescence PCR reaction system and a SYBR Green I real-time fluorescence PCR amplification program. The invention has the beneficial effects that three types of viruses, namely the porcine rabies virus, the porcine parvovirus and the porcine circovirus type 2, can be simultaneously and effectively diagnosed and detected; non-specific swine fever virus, porcine reproductive and respiratory syndrome virus and swine influenza virus can not be detected; and the invention is beneficial to identification and diagnosis of the breeding disorder virus of a pregnant swine, and has better sensitivity, repeatability and stability.

Description

technical field [0001] The invention relates to a real-time fluorescent PCR detection method, in particular to primers and a method for multiplex SYBR Green I real-time fluorescent PCR detection of porcine pseudorabies virus, porcine parvovirus and porcine circovirus type 2. Background technique [0002] Porcine pseudorabies virus (PRV), porcine parvovirus (PPV), and porcine circovirus type 2 (PCV2) can cause reproductive disorders in pregnant sows to varying degrees, causing sow abortion, embryonic death, fetal malformation, and fetal mummification Infertility and infertility, etc., have the characteristics of fast transmission, high embryonic lethality, wide epidemic range, many transmission routes, and stubborn pathogens in pig herds, which cause huge losses to the pig industry every year. And often mixed infection, relying on neutralization test and virus isolation to confirm the diagnosis is likely to cause false negatives, and the operation is cumbersome and takes a lo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64C12R1/93
Inventor 魏战勇李明凤陈红英党玉丽胡慧崔保安郭显坡
Owner HENAN AGRICULTURAL UNIVERSITY
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