Preparation and preliminary application of colloidal gold from Listeria monocytogenes llo

Abstract

The invention provides the preparation and preliminary application of Listeria monocytogenes LLO colloidal gold, anti-Listeria monocytogenes monoclonal antibody hybridoma cell line, and solves the problem that reliable LM monoclonal antibodies are not easy to obtain. Multiple passages can stably secrete monoclonal antibodies. The monoclonal antibodies secreted by it are used in the detection of Listeria monocytogenes in food by colloidal gold method, with good sensitivity, specificity, stability and repeatability Etc. Pathogens can also be detected at very low concentrations, further improving the detection sensitivity and detection rate, and strictly preventing missed and false detections.

Description

technical field [0001] The invention belongs to the field of biotechnology, specifically the preparation and preliminary application of Listeria monocytogenes LLO colloidal gold. Background technique [0002] Listeria monocytogenes (Listeriamonocytogenes, LM) is one of the most common food-borne pathogens, and the WHO listed it as one of the four major food-borne bacteria in the 1990s along with Escherichia coli O157:H7, Salmonella and Staphylococcus aureus. Origin pathogenic bacteria. LM is the most pathogenic bacterium in the genus Listeria, and it is also the only typical intracellular parasite that is pathogenic to humans. It can pass through the three barriers of the host and cause serious zoonotic infectious diseases, such as Inflammation, meningitis, sepsis, miscarriage, etc. Pregnant women, newborns, the elderly and immunocompromised persons have a fatality rate of 20-30% after infection with the bacteria, and neonatal and other immunocompromised persons have a fat...

AI Technical Summary

Problems solved by technology

At present, the detection of LM in my country mainly adopts traditional separation culture detection and identification, PCR, ELISA and automatic identification system and other detection technologies. Traditional separation culture and biochemical identification generally take 3-7 days, the detection cycle is long, and the operation is cumbersome; PCR technology needs to extract nucleic acid and then amplify. The DNA extraction process is easily affected by exogenous nucleic acid. At the same time, impurities and bacteria in food will inhibit PCR amplification, resulting in false negatives. There is no LM in the domestic market. ELISA kit, so LM’s ELISA test needs to buy imported kits, which undoubtedly increases the cost of testing and limits the wide application of ELISA; the price of automatic identification system is high, and it is difficult to popularize in grassroots laboratories

Moreover, there is a common problem in the above tests - in the actual measurement of the actual samples, the food needs to be pre-cultured in order to increase the number of LM through the process of enrichment, but this will undoubtedly prolong the detection time and is not suitable for food-borne diseases. Rapid detection of pathogenic bacteria

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