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A multiplex PCR detection primer set and kit for quickly distinguishing PCV2, PCV3 and Mhp

A detection kit and detection primer technology, applied in the field of zoology and molecular biology, can solve the problems of economic loss, difficult diagnosis, increase the difficulty of swine disease prevention and control and diagnosis, and achieve the effect of easy operation and rapid detection

Inactive Publication Date: 2019-01-11
GUANGXI VETERINARY RES INST
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  • Claims
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AI Technical Summary

Problems solved by technology

Mycoplasma hyopneumoniae can significantly reduce the growth rate and feed conversion rate of infected animals, causing large economic losses
At the same time, it damages the body immunity of infected animals and forms a systemic immunosuppression, which makes infected animals more susceptible to other pathogens, greatly increasing the difficulty of current pig disease prevention and diagnosis.
[0005] PCV2, PCV3, and Mhp can cause respiratory diseases in pigs, and all three are immunosuppressive diseases. After infection, they can be infected with a variety of viral or bacterial diseases, making the epidemic more serious and difficult to diagnose. The pig industry brings even greater economic losses

Method used

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  • A multiplex PCR detection primer set and kit for quickly distinguishing PCV2, PCV3 and Mhp
  • A multiplex PCR detection primer set and kit for quickly distinguishing PCV2, PCV3 and Mhp
  • A multiplex PCR detection primer set and kit for quickly distinguishing PCV2, PCV3 and Mhp

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Experimental program
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Embodiment

[0035] 1 Materials and methods

[0036] 1.1 Viruses and clinical samples

[0037] Porcine parvovirus (PPV), porcine pseudorabies virus (PRV), porcine bocavirus (PBoV), porcine leukovirus (TTV), Escherichia coli, Staphylococcus aureus, Haemophilus parasuis, Actinobacillus pleuropneumoniae Virus strains or bacterial strains are preserved and provided by this laboratory. Clinical samples were collected from dying pigs suspected to be infected with porcine circovirus and mycoplasma hyopneumoniae, dead pigs, or major organs (lymph nodes, spleen, lungs, kidneys, etc.) -1.0g, 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 000r / min for 5min, collect the supernatant, transfer to a 1.5mL centrifuge tube and number for DNA extraction or store at -20°C for later use.

[0038] 1.2 Main reagents

[0039] pMD18-T Vector, Bam HI, Hin dⅢ, DL...

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Abstract

The invention discloses a multiplex PCR detection primer set and a kit for quickly distinguishing PCV2, PCV3 and Mhp, belonging to the technical field of animal diseases and molecular biology. The kitcomprises the primer sequence shown in SEQ ID1-6 and also includes 10*Multi HoeStart Buffer, Super Pure dNTPs, Multi HoeStart DNA Polymerase, template and sterilized water. The multiplex PCR detection kit of PCV2, PCV3 and Mhp provided by the invention adopts multiplex PCR amplification, which is one-time detection of three kinds of pig respiratory tract diseases. The detection method is easy tooperate, convenient and quick, and can achieve the purpose of rapid detection.

Description

technical field [0001] The invention belongs to the technical field of zoology and molecular biology, and specifically relates to a detection primer set, a detection kit and application of multiple PCR for PCV2, PCV3 and Mhp. 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 is the main pathogen that causes multisystemic wasting syndrome (PMWS) in weaned piglets. The diseases are closely related, both porcine circovirus-associated disease (PCVAD). Clinically, PCV2 is often co-infected with pathogens such as porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV) and mycoplasma hyopneumoniae (MHp). At the s...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2537/143
Inventor 刘磊段群棚周英宁赵武李斌何颖秦毅斌卢冰霞陈忠伟张宁梁家幸苏乾莲蓝常力冯建远蒋冬福卢敬专郭旋
Owner GUANGXI VETERINARY RES INST
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