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Double real-time fluorescence quantification PCR (polymerase chain reaction) detection primer for detection of porcine parvovirus type 2 and porcine parvovirus type 3

A real-time fluorescence quantitative, parvovirus technology, applied in the determination/inspection of microorganisms, microorganisms, recombinant DNA technology, etc., to achieve the effect of strong specificity and high sensitivity

Inactive Publication Date: 2015-04-01
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, there is no relevant research report on the detection of porcine parvovirus type 2 and type 3 (PPV2 and PPV3) based on SYBR I dual real-time fluorescent PCR. The establishment of the present invention can fill the gap in the related field

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  • Double real-time fluorescence quantification PCR (polymerase chain reaction) detection primer for detection of porcine parvovirus type 2 and porcine parvovirus type 3
  • Double real-time fluorescence quantification PCR (polymerase chain reaction) detection primer for detection of porcine parvovirus type 2 and porcine parvovirus type 3
  • Double real-time fluorescence quantification PCR (polymerase chain reaction) detection primer for detection of porcine parvovirus type 2 and porcine parvovirus type 3

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Experimental program
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Embodiment Construction

[0016] 1.1 Materials

[0017] 1.1.1 Virus strains and bacterial strains

[0018] PPV2 and PPV3 positive DNA was identified and preserved by our hospital.

[0019] 1.1.2 Main instruments and reagents

[0020] SYBR? Primemix Ex Taq TM Ⅱ (Tli RNaseH Plus), pMD18-T vector, DL500 DNA Marker, EASY Dilution were purchased from Treasure Bioengineering (Dalian) Co., Ltd.; rapid DNA extraction and detection kit (KG203), gel recovery kit , plasmid mini-extraction kit, and Taq PCR Mastermix (KT201) were purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.; fluorescent quantitative PCR tubes were purchased from Axygen.

[0021] 1.2 Method

[0022] 1.2.1 Primer design and synthesis

[0023] Referring to the genome characteristics of PPV2 and PPV3 registered in GenBank, DNAStar software was used for homology analysis and comparison, and conserved and specific nucleotide regions were selected, and primers were designed using the primer design software Oligo7.0 software (see T...

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Abstract

The invention provides a double real-time fluorescence quantification PCR (polymerase chain reaction) detection primer and method for detection of porcine parvovirus type 2 (PPV2) and porcine parvovirus type 3 (PPV3), wherein the primer sequence of the PPV2 is as follows: an upstream primer P1 is GAAGCGGGGAAGATACTC, and a downstream primer P2 is GTCCTTAGCTCTCCACAG; the primer sequence of the PPV3 is as follows: an upstream primer P3 is GAGGGAATTTAGCAACTG, and a downstream primer PR is TCTGGTCTTCCTGATATC. Research reports related to detection of porcine parvovirus type 2 (PPV2) and porcine parvovirus type 3 (PPV3) based on SYBRI double real-time fluorescence PCR are not disclosed at present, and the development of the invention can fill up blanks in related fields.

Description

technical field [0001] The invention relates to dual real-time fluorescent quantitative PCR detection primers and methods for porcine parvovirus type 2 and type 3 (PPV2 and PPV3), in particular to primers and a method for multiple SYBR Green I real-time fluorescent quantitative PCR methods for PPV2 and PPV3. Background technique [0002] With the deepening of viral metagenomics research, there are more and more reports on the discovery of new viruses. In recent years, there have been more and more reports of DNA viruses in pigs. In addition to the classic porcine parvovirus (PPV) and porcine circovirus (porcine circovirus type Ⅰ and Ⅱ, PCV1 and PCV2), porcine parvovirus 2 has also been reported. Porcine parvovirus type 2 (PPV2), porcine parvovirus type 3 (PPV3), porcine parvovirus type 4 (PPV4) and various subtypes of porcine bocavirus , PBoV). [References: Xiao CT, Giménez-Lirola LG, Jiang YH, Halbur PG, Opriessnig T. Characterization of a novel porcine parvovirus tentati...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/70C12Q2537/143C12Q2527/107C12Q2561/113
Inventor 李家玉张奇杨晓燕
Owner FUJIAN AGRI & FORESTRY UNIV