Method for making recombinant antigen pp 65 envelope enzyme-linked reaction plate and ELISA test kit

Abstract

The present invention relates to a new method for making recombinant antigen PP65 coated enzyme labeled reaction plate and ELISA kit. Its key technique mainly liesin that using gene engineering method to prepare specific recombinant antigen-HCMV pp65, after the antigen is exracted and purified, using the coating plate as important component in the kit, besides, after the anti-human IgM monoclone antibody prepared by utilizing mouse-mouse hybridome technique is labeled by using horseradish peroxidase, it can be used for assembly of kit. Said method capable of creating sensitive quick ELISA antibody to trap HCMV specific IgM antibody in serum has high specificity and high stability, and said kit mainly can be used in laboratory research and clinical diagnosis.

Description

technical field [0001] The invention relates to a novel biological preparation HCMVpp65 recombinant protein, which is an ELISA detection kit for detecting HCMV-IgM. Background technique [0002] Human cytomegalovirus (Human Cytomegalovirus, HCMV) belongs to the human herpesvirus beta subfamily virus, the infection in the population is very common, showing a worldwide distribution. Serological epidemiological surveys show that 40-100% of adults have HCMV antibodies; different countries, different economic conditions, different groups of people, different age groups, HCMV infection rates are different, and its prevalence has no seasonal tendency. In Asia and Africa, 90% of the population is infected, and most of them have antibodies in their adolescence, indicating that the age of infection is earlier, while the infection occurs later in Western countries; the infection rate of people with low economic income is higher than that of people with higher economic income. HCMV is ...

AI Technical Summary

Problems solved by technology

At present, the quality of various immunodiagnostic kits for HCMV in the market is uneven, there is no uniform quality standard, and the production and application are out of order, which has caused serious obstacles to the prevention, diagnosis and treatment of HCMV

At present, although HCMV in vitro diagnostic reagents are on the market at home and abroad, the foreign products are expensive and not suitable for my country's national conditions.

Domestic kits mostly use cell-cultured whole virus crude antigens to detect specific IgM in serum, the virus yield is low, and the purity of the obtained virus antigens is low, and its antigenicity may cross with other herpes viruses, which affects the accuracy of the test results ; These kits have disadvantages such as poor stability and repeatability, low sensitivity and specificity

In addition, investigations and comparisons have confirmed that there are large quality differences among the products produced by various manufacturers, and the minimum detection amount can vary by 15 to 30 times.

The main reason for the quality differences of domestic testing reagents may be related to the manufacturer's indifference to quality awareness and lack of quality control standards