Dual SYBR Green I real-time fluorescence PCR detection primer and method for porcine parvovirus and porcine circovirus type 2
A porcine circovirus and parvovirus technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fluorescence/phosphorescence, etc., can solve the problems of inability to quantify, false negative, spread, etc., and achieve good specificity
Inactive Publication Date: 2011-05-25
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology
However, due to the large difference between different primers and the influence of various factors during PCR amplification, false negative results are often prone to occur during diagnosis, and cannot be quantified; especially for recessively infected animals, often missed detection, resulting in Epidemic and spread of disease
Although the TaqMan probe method established by PRV and PPV can be qualitative and quantitative, the cost is high, which is not conducive to popularization and application
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[0026] The sequences of porcine parvovirus and porcine circovirus type 2 dual SYBR Green I real-time fluorescent PCR detection primers are as follows:
[0027] The sequence of the porcine circovirus type 2 primer is as follows:
[0028] Upstream primer P1: 5′-TAA CTA CTC CTC CCG CCA TAC-3′;
[0029] Downstream primer P2: 5′-GCC TAC GTG GTC TAC ATT TCC-3′;
[0030] The sequences of porcine parvovirus primers are as follows:
[0031] Upstream primer P3: 5′-TGG GAG GGC TTG GTT AGA-3′;
[0032] Downstream primer P4: 5'-TGG TGG TGA GGT TGC TGA T-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 parvovirus 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 parvovirus 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 according to 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 parvovirus and the porcine circovirus type 2 can be detected simultaneously, and porcine pseudorabies virus, 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 specificity, repeatability and sensitivity.
Description
technical field [0001] The invention relates to a real-time fluorescent PCR detection method, in particular to a double SYBR Green I real-time fluorescent PCR detection method for porcine parvovirus and porcine circovirus type 2. Background technique [0002] Porcine parvovirus (PPV) is a viral infectious disease mainly characterized by sow reproductive disorders caused by porcine parvovirus infection. It is characterized in that when sows are infected with the virus in the early pregnancy, the embryo or fetus is attacked through the placenta. It can cause miscarriage, embryonic death, fetal malformation, fetal mummification and infertility in sows. It can also cause dermatitis and diarrhea in piglets, but other types of pigs have no obvious clinical symptoms after infection. PPV has a very high detection rate in pigs, and the positive rate of serum antibodies in pigs reaches 50% to 80%. It is one of the main viral reproductive disorders that plague the development of the pi...
Claims
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Login to View More IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64C12R1/93
Inventor 崔保安魏战勇陈红英李明凤党玉丽郭显坡
Owner HENAN AGRICULTURAL UNIVERSITY
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