Lipoteichoic acid enzyme-linked immunosorbent assay (ELISA) detection kit and preparative technique thereof

Abstract

The invention relates to preparation of lipoteichoic acid monoclonal antibodies and an e-linked immunosorbent assay (ELISA) detection kit manufactured by using the preparation. An ELISA detection reagent is a high-technology product in the field of biological medicines, and is an in-vitro diagnostic reagent. Following a search, the ELISA detection reagent fills the domestic blank, the current situation of detecting staphylococcus aureus by using a time-wasting and labor-wasting blood culture method is changed, an enzyme linked immunosorbent assay which is high in sensitivity and specificity is adopted, and detection time is shortened. Balb / c female mice are immuned by lipoteichoic acid antigens extracted from cell walls of the staphylococcus aureus, then monoclonal antibodies L are obtained and are IgG immune globulin after being detected, and the valence of the antibodies is 1*106. The monoclonal antibodies are purified by a saturated ammonium sulfate precipitation and affinity chromatography technology. Lipoteichoic acids serve as coating antigens, the obtained monoclonal antibodies serve as enzyme-labeled antibodies, and then a competition-method ELISA kit is manufactured and is used for diagnosing staphylococcus aureus infection which serves as one of references for clinical diagnosis.

Description

Technical field: [0001] The invention relates to a lipoteichoic acid ELISA detection kit and its preparation technology, which are used for the rapid detection of clinical Staphylococcus aureus infection and belong to the field of biomedicine. Background technique: [0002] With the aging of the population, the widespread development of organ transplant operations, the use of immunosuppressants, and the influence of social factors, the incidence of Gram-positive bacterial infections has increased, and the pathogenic bacteria of bacteremia accounted for Gram-positive bacteria. 59.6%, of which Staphylococcus accounts for 38.6%. At present, automated blood culture systems have been widely used in clinical microbiology laboratories. Under normal circumstances, when a positive alarm bottle appears, the bacteria in the bottle are first transferred to the blood plate, and then the identification and drug sensitivity test are performed. The result is 48-72 hours after the positive alarm...

AI Technical Summary

Problems solved by technology

At present, the automated blood culture system has been widely used in clinical microbiology laboratories. Usually, when a positive alarm bottle appears, the bacterial solution in the bottle is first transferred to a blood plate, and then identification and drug sensitivity tests are performed. The result is that 48-72 hours after the positive alarm, this method is time-consuming, laborious and inefficient, and is very unfavorable to clinical anti-infection treatment. Therefore, a rapid, accurate and sensitive detection method is urgently needed for clinical diagnosis of Gram-positive bacterial infection