Method of quickly detecting content of cell infective virions in circovirus

A virus particle and circovirus technology, applied in the field of biological genes, can solve the problems of long detection cycle, large human error, and inability to distinguish infectious virus particles, so as to shorten detection time, improve accuracy and repeatability Effect

Inactive Publication Date: 2016-02-17
SICHUAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECHN CENT +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method has a long detection period, complicated operation steps, and large human error.
PCR/qPCR methods cannot distinguish between infectious and

Method used

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  • Method of quickly detecting content of cell infective virions in circovirus
  • Method of quickly detecting content of cell infective virions in circovirus
  • Method of quickly detecting content of cell infective virions in circovirus

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[0024] The present invention will be further described below in conjunction with the accompanying drawings and specific examples, but it is not intended to limit the present invention.

[0025] 1. Test materials

[0026] 1.1 Main test equipment

[0027] Bole C-1000 PCR instrument, Bole CFX96 fluorescence quantitative PCR instrument, ultra-low temperature refrigerator, ultraviolet spectrophotometer, high-speed centrifuge, biological safety cabinet, electrophoresis instrument, chemiluminescence gel imaging system

[0028] 1.2 Test materials

[0029] The viral DNA / RNA magnetic bead extraction kit was purchased from Chengdu Tuoyang Technology Co., Ltd.; the fluorescence quantitative PCR detection reagent 2×SsoFastTM Probes Supermix was purchased from Bó Lè; DNA molecular weight standards and agarose were purchased from Bao Bioengineering (Dalian) Co., Ltd.; Other reagents are of domestic analytical grade. The circovirus vaccine strain was provided by Sichuan Huapai Biotechnology Company. ...

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Abstract

The invention relates to a method of quickly and quantitatively detecting the content of cell infective virions in circovirus and belongs to the technical field of virus detection. The method comprises construction of a circovirus standard plasmid and establishment of a detection standard curve, a target cell is infected with a virus to be detected and a control virus under the same conditions, viral genomes infected with the virus to be detected and the control virus are extracted separately, and the relative contents of the virus to be detected and the control virus are obtained by fluorescent quantitative PCR (polymerase chain reaction) detection. A cell adsorption process is added before the step of fluorescent quantitative PCR detection, and the content of circovirus that is cell infective is quickly measured.

Description

technical field [0001] The invention relates to the field of biological genes, in particular to a method for detecting the content of infectious circular virus particles. Background technique [0002] Porcine circovirus (Porcine circovirus, PCV) is one of the smallest known mammalian viruses. It has two subtypes, PCV1 and PCV2. Among them, PCV1 is not pathogenic to pigs, and PCV2 is the cause of multisystemic wasting syndrome in weaned piglets. The main pathogen is one of the main diseases threatening the global pig industry. Porcine circovirus can replicate in porcine macrophages, porcine monocytes and other cells. In vitro, porcine circovirus can be propagated by PK-15 and IBRS-2 cells, but the virus does not produce cytopathic changes during the passage. Cannot grow on Vero cells, BHK-21 cells, ST cells, etc. At present, the detection methods of porcine circovirus mainly include virus isolation, indirect immunofluorescence test, immunoperoxidase monolayer test, PCR / flu...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
Inventor 林华陈世界杨苗肖艳严玉宝胡娟赵姗薛昌华
Owner SICHUAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECHN CENT
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