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Quantitative detection method for porcine circovirus type II virus-like particles

A quantitative detection method and technology for porcine circovirus, applied in the biological field, can solve the problems of non-commercialization, inability to maintain the conformation of VLPs, and interference with the detection signal of PCV2 VLPs.

Active Publication Date: 2019-01-11
FUDAN UNIV
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Problems solved by technology

[0006] However, the existing HPLC detection methods applied to PCV2 VLPs are difficult to apply to the quantitative detection of PCV2 VLPs, and domestic and foreign literature and patent searches have not retrieved relevant reports. Molecular complexes, conventional reversed-phase HPLC and normal-phase HPLC cannot maintain the conformation of VLPs during detection; 2. After the yeast cells are broken, a large amount of dissolved substances such as proteins, nucleic acids and polysaccharides are released, some of which will interfere with the detection signal of PCV2VLPs, It may even interact with PCV2VLPs and affect the detection effect; 3. Due to the difficulty in purification and quantification of PCV2VLPs, currently there are no commercial standards used to make standard curves

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  • Quantitative detection method for porcine circovirus type II virus-like particles
  • Quantitative detection method for porcine circovirus type II virus-like particles
  • Quantitative detection method for porcine circovirus type II virus-like particles

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Embodiment Construction

[0022] The quantitative detection method of porcine circovirus type 2 virus-like particle (hereinafter referred to as virus-like particle) that the present invention adopts mainly comprises the following steps:

[0023] Step S1, diluting the sample to be tested and performing filtration clarification or centrifugation clarification;

[0024] Step S2, take the porcine circovirus type 2 virus-like particle standard product whose protein content has been determined, and perform serial dilution, and perform HPLC detection on the diluted serial standard product, and establish a curve with the concentration as the abscissa and the peak height as the ordinate. regression equation;

[0025] Step S3, performing HPLC detection on the diluted sample to be tested to obtain the corresponding sample peak height;

[0026] In step S4, the concentration of porcine circovirus type 2 virus-like particles in the sample to be tested is calculated according to the regression equation and the peak ...

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Abstract

The invention aims to provide a quantitative detection method for porcine circovirus type II virus-like particles simple to operate, and high in sensitivity, repeatability and applicability. The method comprises the following steps: S1, diluting to-be-tested samples and filtering and clarifying or centrifugalizing and clarifying the samples; S2, serially diluting the porcine circovirus type II virus-like particle standard substances with determined protein contents, and carrying out HPLC detection on the diluted serial standard substances to establish a regression equation which takes concentration as a horizontal ordinate and peak height as a vertical ordinate; S3, carrying out HPLC detection on the diluted to-be-tested samples to obtain corresponding peak heights of the samples; and S4,calculating the concentrations of the porcine circovirus type II virus-like particles in the to-be-tested samples according to the regression equation and the peak heights of the samples, wherein chromatographic columns used in HPLC in the S2 and S3 are gel chromatographic columns, and detectors are ultraviolet detectors, the detection wavelengths of which are 280 nm.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a quantitative detection method of virus-like particles, in particular to a quantitative detection method of porcine circovirus type 2 virus-like particles. Background technique [0002] Porcine circovirus (porcine circovirus, PCV) belongs to Circoviridae (Circoviridae) genus Circovirus (Circovirus), is the smallest virus found so far, its smallest particle diameter is only 17nm, DNA genome length 1.7kb, virion It is an icosahedral symmetrical, non-enveloped, single-stranded circular DNA virus. There are two known serotypes of PCV, non-pathogenic PCV1 and pathogenic PCV2. PCV2 is the main pathogen causing postweaning multisystemic wasting syndrome (PostweaningMultisystemic Wasting Syndrome, PMWS), mixed infection with classical swine fever virus (CSFV) or porcine reproductive and respiratory syndrome virus (PRRSV) can enhance the severity of the disease, It can cause a mortality rate...

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 吕红周峻岗段进坤杨德强
Owner FUDAN UNIV
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