Duplex PCR (polymerase chain reaction) detection primer and kit for quickly distinguishing porcine circoviruses type 2 and type 3

A technology of porcine circovirus and detection kit, applied in the field of animal virology and molecular biology, to achieve high sensitivity and easy detection method

Inactive Publication Date: 2018-09-14
GUANGXI VETERINARY RES INST
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  • Abstract
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  • Claims
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Problems solved by technology

As a new animal pathogen, PCV3's pathogenicity to pigs and its role in PCVAD, as

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  • Duplex PCR (polymerase chain reaction) detection primer and kit for quickly distinguishing porcine circoviruses type 2 and type 3
  • Duplex PCR (polymerase chain reaction) detection primer and kit for quickly distinguishing porcine circoviruses type 2 and type 3
  • Duplex PCR (polymerase chain reaction) detection primer and kit for quickly distinguishing porcine circoviruses type 2 and type 3

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Embodiment Construction

[0022] 1 Materials and methods

[0023] 1.1 Viruses and clinical samples

[0024] Porcine parvovirus (PPV), porcine pseudorabies virus (PRV), Escherichia coli, Staphylococcus aureus, Haemophilus para, and Actinobacillus pleuropneumoniae strains or strains are all provided by the laboratory. Clinical samples were collected from the main organs (lymph nodes, spleen, lungs, kidneys, etc.) of dying pigs suspected of being infected with circovirus in pig farms in various places in Guangxi, dead pigs, or pigs that were slaughtered. About 0.5g to 1.0g of tissue samples were cut , placed in a sterilized mortar, fully ground into a homogenate, added sterilized 1mol / LPBS solution to make a 1:5 emulsion suspension, and repeated freezing and thawing 3 times. Centrifuge at 10 000 r / min for 5 min, collect the supernatant, transfer to a 1.5 mL centrifuge tube and number for DNA extraction or store at -20°C for later use.

[0025] 1.2 Main reagents

[0026] pMD18-T Vector, Bam HI, Hin ...

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Abstract

The invention belongs to the technical field of animal virology and molecular biology and discloses a duplex PCR (polymerase chain reaction) detection primer and kit for quickly distinguishing porcinecircoviruses type 2 and type 3. The kit comprises primer sequences shown as SEQ ID1-4, 2*F8 Fastlong PCR MasterMix, a cDNA template and sterile water. The duplex PCR detection kit for quickly distinguishing the porcine circoviruses type 2 and type 3 has advantages that by duplex PCR amplification, two types of DNA viruses similar in lesion can be detected in one time, the total detection time iscontrolled to be about two hours, a detection method is easy in operation, convenient and quick, and quick detection can be realized.

Description

technical field [0001] The invention belongs to the technical field of animal virology and molecular biology, and specifically relates to detection primers, detection kits and applications for porcine circovirus type 2 and porcine circovirus type 3 one-step double PCR. Background technique [0002] Porcine circovirus (PCV) is a covalently closed, single-stranded circular DNA virus and is the smallest animal virus discovered so far. Before 2015, only two genotypes of PCV1 and PCV2 were found worldwide. Among them, PCV1 is considered to be a PK-15 pollutant in pig kidney cells, but it does not produce cytopathic effects and is not pathogenic to pigs. PCV2 can cause porcine circovirus-associated diseases (PCVAD), mainly weaned piglet multisystem wasting syndrome (PMWS), porcine dermatitis and nephrotic syndrome (PDNS), porcine respiratory disease syndrome (PRDC), reproductive disorders, hyperplasia Necrotizing pneumonia (PNP) and enteritis, etc. Clinically, PCV2 is often co-...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/686C12Q1/701C12Q2600/16C12Q2537/143
Inventor 卢冰霞秦毅斌何颖赵武刘磊段群棚陈忠伟李斌梁家幸闭炳芬苏乾莲蒋冬福周英宁侯韶毅李茂宁
Owner GUANGXI VETERINARY RES INST
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