Preparation method of measles virus lysate as coating antigen of ELISA kit

Abstract

The invention discloses a preparation method of a measles virus lysate as a coating antigen of an ELISA kit. The preparation method comprises culturing an attenuated measles virus strain to obtain a measles virus liquid, separating a measles virus concentrated solution through ultrafiltration or other methods, adding a measles virus antigen protective fluid into the measles virus concentrated solution to obtain a measles virus stock solution having a certain concentration, preserving the measles virus stock solution for next use, carrying out pyrolysis treatment, adjusting the concentration of the measles virus stock solution through a sample dilution solution to obtain a measles virus lysate directly as a coating antigen, adjusting pH of the coating and fixing solutions and ion strength, and selecting an appropriate buffer pair. The preparation method improves detection sensitivity and specificity and satisfies clinical detection demands.

Description

technical field [0001] The invention relates to the field of clinical experiments, in particular to a method for preparing a measles virus lysate used as an enzyme-linked kit for coating antigens. Background technique [0002] Morbillivirus belongs to Family Paramyxoviridae (Family Paramyxoviridae) and Morbillivirus (Genus Morbillivirus). It is a negative-strand RNA virus. It is different from other paramucosal viruses in that it has no special neuraminidase and is spherical in shape with a diameter of only 100-250nm. , Helical symmetry, enveloped, containing hemagglutinin on the envelope. The total length of the genome is about 16kb. The genome has six genes, N, P, M, F, H, and L, which encode six structural and functional proteins: nucleoprotein NP, phosphorylated protein P, membrane protein M, fusion protein F (fusion protein, F), hemagglutinin protein H (haemagglutinin protein, H), RNA-dependent RNA polymerase (Large polymerase, L). Their components are all glycoprotein...

AI Technical Summary

Problems solved by technology

The cost of using recombinant antigens to replace natural whole virus antigens is higher, and the structural and functional protein fragments of recombinant measles virus expressed by genetic engineering techniques often have incomplete epitope components

In addition, the recombinant antigen obtained by prokaryotic expression, separation and purification of E. coli contains other miscellaneous proteins of E. coli. However, in people who have been infected by E. coli, the anti-E. coli antibodies in the serum can react with these miscellaneous proteins and cause false positives. These miscellaneous proteins are likely to affect the detection results