Method for detecting foot-and-mouth disease antigen 146S content with sucrose density gradient ultraviolet light quantitative method

Abstract

The invention provides a method for detecting foot-and-mouth disease antigen 146S content with a sucrose density gradient ultraviolet light quantitative method; the method comprises the following steps: sucrose density gradient ultra centrifugation is carried out to virus concentration liquid to be detected, and then OD259nm value of all fractions is continuously detected by an ultraviolet light spectrophotometer; next, peak area of OD value of a sample to be detected is calculated, and the 146S content in the virus liquid is calculated out according to the formula (FR*PA*FSD*1000) / (S*PL*E*W),wherein FR is flow rate of sucrose which passes through a sample cell; PA is peak area; S is speed, FSD is an absorbance unit, PL is optical path length of the flowing sample cell, W is original weight or volume of a sample which is added on the gradient, E is correction coefficient which is optional positive number. The method has better repetitiveness and applicability, and can be used for accurate and quantitative detection of 146S relative content of various foot-and-mouth viruses.

Description

technical field [0001] The invention relates to a method for quantitatively detecting the content of the foot-and-mouth disease antigen 146S, in particular to a method for detecting the content of the foot-and-mouth disease antigen 146S by a sucrose density gradient ultraviolet light quantitative method. Background technique [0002] The FMD antigen quantitative detection techniques that have been applied or reported at present mainly include: 1. The half-lethal dose (LD50) of suckling mice is used to measure the FMD virus antigen content. Dilution, each titer subcutaneously injects 4 suckling mice by the amount neck back of every 0.2ml, finally calculates virus antigen content according to suckling mouse morbidity and death situation; For example, make a 10-fold serial dilution of the virus on a 96-well flat-bottom culture plate, the amount of virus solution in each well is 50ml, then add 100ml of cell suspension, and culture in a 37°C, 5% carbon dioxide incubator for 72 ho...

AI Technical Summary

Problems solved by technology

Method 1 and method 2 are for the detection of viral nucleic acid, which can not only detect the nucleic acid of intact virions, but also the nucleic acid of 146S cleavage products, which cannot reflect the immunogen of the virus; method 3 is interfered by pre-complement and anti-complement factors, The accuracy and sensitivity of the detection are restricted, and it cannot distinguish 12S and 146S well; the main problem of method 4 is that it cannot distinguish 12S and 146S well; although method 5 can distinguish 12S and 146S well, However, for different types of viruses, this method must screen out their own monoclonal antibodies, and establish their own independent ELISA detection and calculation systems

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