Dual SYBR Green I real-time fluorescence PCR detection primer and method for porcine parvovirus and porcine pseudorabies virus
A porcine pseudorabies virus and parvovirus technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fluorescence/phosphorescence, etc., can solve the problems of mixed infection, inability to distinguish and determine the infection of a certain disease, and achieve good specificity sexual effect
Inactive Publication Date: 2012-08-29
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology
[0004] The technical problem to be solved in the present invention is to provide a kind of porcine parvovirus and porcine pseudorabies virus that can not distinguish and determine the infection of a certain disease clinically by observing clinical symptoms and pathological changes, and sometimes mixed infection. Duplex SYBR Green I real-time fluorescent PCR detection method for detecting porcine parvovirus and porcine pseudorabies virus
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[0030] The sequence of the primers for detection of porcine parvovirus and porcine pseudorabies virus double SYBR Green I real-time fluorescent PCR is as follows:
[0031] The sequence of porcine parvovirus primers is as follows:
[0032] Upstream primer P1: 5'-TAA TTT AAT CAA CAG GCA CCA C-3';
[0033] Downstream primer P2: 5'-TTC TGT ATC AAG TTC TTT ATC CC-3';
[0034] The primer sequence of porcine pseudorabies virus is as follows:
[0035] Upstream primer P3: 5'-CGT GGA ACG AGC CCT TCA G-3';
[0036] Downstream primer P4: 5'-AGA GCG GGT TGG CGA TGT-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 pseudorabies virus. 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 pseudorabies virus 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 parvovirus and the porcine pseudorabies virus can be detected simultaneously, and porcine circovirus type 2 (PCV2), 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, and contribute to the identification and the diagnosis of pregnant sow reproductive disturbance virus disease.
Description
Technical field [0001] The invention relates to a real-time fluorescent PCR detection method, in particular to a real-time fluorescent PCR detection method for the double SYBR Green I of porcine parvovirus and porcine pseudorabies virus. Background technique [0002] Porcine parvovirus (PPV) and porcine pseudorabies virus (PRV) can infect sows, causing sow abortion, embryonic death, fetal malformation, fetal mummification, and infertility. It can also cause dermatitis and diarrhea in piglets. It can also infect piglets, cause piglet death or form stiff pigs, causing huge economic losses to the pig industry. Clinically, it is impossible to distinguish and determine the infection of a certain disease by observing the clinical symptoms and pathological changes, and there are mixed infections from time to time, which makes clinical detection and control of these two diseases very difficult. [0003] Fluorescence quantitative PCR detection technology has only been successfully develope...
Claims
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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64C12R1/93
Inventor 魏战勇陈红英胡慧李明凤崔保安郭显坡
Owner HENAN AGRICULTURAL UNIVERSITY