Fourfold real-time fluorescent PCR identifying and detecting method for porcine circoviruses 1 to 4

A technology of porcine circovirus and real-time fluorescence, which is applied in the biological field to achieve the effect of improving detection efficiency, high sensitivity and good repeatability

Inactive Publication Date: 2020-10-13
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The detection method of the present invention only needs one detection to determine whet...

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  • Fourfold real-time fluorescent PCR identifying and detecting method for porcine circoviruses 1 to 4
  • Fourfold real-time fluorescent PCR identifying and detecting method for porcine circoviruses 1 to 4
  • Fourfold real-time fluorescent PCR identifying and detecting method for porcine circoviruses 1 to 4

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Embodiment 1

[0036] Establishment and evaluation of quadruple real-time fluorescent PCR differential detection method for porcine circovirus types 1 to 4 (PCV1-4)

[0037] 1.1 Design of primers and probes

[0038] According to the whole genome of PCV strains obtained from the GenBank database, the sequence alignment software DNAMAN was used for multiple sequence alignment analysis, and four specific TaqMans for identifying PCV1, PCV2, PCV3, and PCV4 were designed based on the highly conserved regions in the ORF1 and ORF2 genes. probe and 4 pairs of primers. The 5' ends of the probes were labeled with different fluorophores (FAM, HEX, ROX and TAMRA). The primers and probes in PCV quadruple real-time fluorescent PCR are shown in Table 1-1.

[0039] Table 1-1. Primers and probes used to construct PCV quadruple real-time fluorescent PCR differential detection method

[0040]

[0041]

[0042] *The positions of corresponding primers and probes in the genome are determined according to PC...

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Abstract

The invention relates to a fourfold real-time fluorescent PCR identifying and detecting method for porcine circoviruses 1 to 4, and belongs to the technical field of biology. In accordance with ORF1 and ORF2 genes of different genotypes of PCV, specific primers and probe sequences, capable of identifying PCV1, PCV2, PCV3 and PCV4, and a probe of the PCV1-4 strains labeled with different fluorescein can be designed; each probe can be labeled with any fluorescein, and fluorescein for labeling the four probes needs to be detected by different detection passages; the primers and probes for detecting the PCV1-4 strains, or the primer and probe for identifying the PCV1-4 strains are placed in the same reaction tube, a fluorescent PCR amplification instrument is used for a real-time fluorescent PCR reaction, and the same sample is detected once, so that identification and detection on the PCV 1-4 strains can be completed.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a quadruple real-time fluorescent PCR detection method, in particular to a quadruple real-time fluorescent PCR identification and detection method for types 1 to 4 of porcine circovirus. Background technique [0002] Porcine circovirus (Porcine circovirus, PCV) belongs to Circoviridae, Circovirus genus. PCV is the smallest circular single-stranded DNA virus known so far. The diameter of the virus particle is about 17nm, without envelope, and it is icosahedral in symmetry. Currently, PCV can be divided into four genotypes (PCV1, PCV2, PCV3, PCV4). In 1974, PCV1 was first discovered from the contaminated porcine kidney cell line PK-15. PCV1 is currently considered non-pathogenic to pigs. In 1998, it was first reported that PCV2 was closely related to multisystem wasting syndrome and porcine dermatitis nephrotic syndrome in weaned piglets. In 2015, PCV3 was first identified in porcine ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2537/143C12Q2561/101
Inventor 陈南华肖延昭朱建中
Owner YANGZHOU UNIV
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