Method for Plaque Titration of Foot-and-Mouth Disease Virus in Suspension Cells

A technology of foot-and-mouth disease virus and suspension cells, which is applied in the field of plaque detection of foot-and-mouth disease virus suspension cells, can solve the problems of large detection errors and achieve the effects of small batch-to-batch errors, reasonable use, and stable production

Active Publication Date: 2012-02-01
JINYUBAOLING BIO PHARMA CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a kind of accurate standardization foot-and-mouth disease virus virulence assay method, to overcome the problem that the existing FMD virus virulence assay method is affected by factors such as living animal individual difference and brings the larger problem of detection error

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  • Method for Plaque Titration of Foot-and-Mouth Disease Virus in Suspension Cells
  • Method for Plaque Titration of Foot-and-Mouth Disease Virus in Suspension Cells
  • Method for Plaque Titration of Foot-and-Mouth Disease Virus in Suspension Cells

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Embodiment Construction

[0025] The present invention will be further described below through specific examples.

[0026] The foot-and-mouth disease virus suspension cell plaque detection method comprises the following steps:

[0027] (1) Agarose saline solution and serum-free medium are mixed according to a volume ratio of 1:1 to obtain an agar solution, wherein the agarose saline solution contains 2% by weight of agarose and 0.85% by weight of NaCl. Heat in medium until completely dissolved (when heating, pay attention to the air permeability of the outlet) and then keep it warm at 40°C; the serum-free medium is the BBSM medium produced by Tsingda Tianyi Company, prepared according to its instructions;

[0028] (2) Add 1ml of the agar solution to the six-hole cell culture plate, shake the cell culture plate, so that the agar solution covers the bottom layer of the cell culture plate to obtain the bottom agar culture plate, place it in a flat position, cool and solidify ;

[0029] (3) BHK-21 suspen...

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Abstract

The invention provides a method for titrating foot-and-mouth disease virus with suspended cell plaques, comprising the following steps: (1) mixing agarose saline solution and serum-free medium to obtain an agar solution; (2) adding the agar solution to cell culture (3) obtain cell suspension; (4) dilute the foot-and-mouth disease venom to be measured to obtain the gradient concentration foot-and-mouth disease venom; (5) obtain the upper layer agar solution and the control containing each gradient concentration foot-and-mouth disease virus (6) the upper layer agar solution containing each gradient concentration of foot-and-mouth disease virus and the upper layer agar solution used for comparison are added to each culture well of the bottom agar culture plate respectively, after solidification, seal with a parafilm, and invert Cultivate in a CO2 incubator to obtain a plaque counting plate; (7) stain with neutral red, and observe the number of plaques in each culture well of the plaque counting plate against the light; (8) calculate the poison price. The detection conditions of the method are uniform, the controllability is strong, and the error between batches is small.

Description

technical field [0001] The invention relates to a method for measuring the valence of foot-and-mouth disease virus (FMDV), in particular to a method for detecting the virus of the foot-and-mouth disease virus in suspended cell plaques. Background technique [0002] At present, the methods for determining the virulence of FMD virus generally adopt two methods, the median infectious dose of cells (TCID50) and the median lethal dose of suckling mice (LD50). The LD50 method needs to use live animals, and the individual differences of live animals will have a great impact on the test results. The TCID50 method needs to prepare monolayer cells in advance, and needs to add serum during the experiment, which makes the detection process limited by the growth state of the cells, and the detection results will also be affected by the added serum. [0003] In addition, suspension cell plaque assay is also an effective method for the determination of virus valence. After the virus infe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12R1/93
Inventor 王伟峰卢永干周劲松刘艳霞
Owner JINYUBAOLING BIO PHARMA CO LTD
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