Quantitative detection method of escherichia coli in soil and assay kit thereof

Abstract

The invention discloses a quantiative detection method of escherichia coli in soil and an assay kit thereof. On the basis of taking related researches as reference, the method performs quantitative detection on pathogenic microorganism in the soil by adopting a method of combining MPN (most probable number) with PCR (polymerase chain reaction). By utilizing advantages of quickness and accuracy ofthe PCR method, biochemical identification, serological identification and other steps of positive results in the traditional MPN method are substituted, so that complex steps in a national standard method are reduced, the detection efficiency is improved, and a pre-enrichment step in the national standard method is adopted at the same time to improve the accuracy of detection targeting pathogenic bacteria; and due to the complexity of soil, the soil contains massive microorganisms, as well as various pathogenic microorganisms of different items, so that the targeting pathogenic bacteria is selectively cultured to improve the detection efficiency in order to quantitatively detect the targeting pathogenic bacteria better and more accurately. By virtue of creative effort of the inventor andscientific test verification, the sensitivity of the escherichia coli quantitative detection method in an experiment of adding pure bacteria in the soil is 25 per gram of soil.

Description

technical field [0001] The invention belongs to the technical field of microorganism quantitative detection, and in particular relates to a method for quantitatively detecting Escherichia coli in soil and a detection kit thereof. Background technique [0002] With the acceleration of urbanization in our country, cities migrate and expand to the edge. The urban-rural fringe is at the junction of urban development and rural construction areas. Soil health and quality issues in the urban-rural fringe are too important to ignore. At present, experts and scholars mainly focus on the indicators of pesticide residues, heavy metal pollution and organic pollution in the research of soil quality, and pay little attention to the biological indicators of pathogenic microorganisms in soil. Pathogenic microorganisms from poultry manure and sewage irrigation soil can remain latent in the soil for a long period of time, and can enter vegetables, eventually affecting human health. In parti...

AI Technical Summary

Problems solved by technology

Among them, the traditional detection method is highly authoritative, but it has the disadvantages of time-consuming, labor-intensive and troublesome operations, and the quantitative detection of pathogenic microorganisms in soil has certain complexity and operational difficulty; with the modern molecular biology With the development of scientific methods, PCR technology is widely used in the qualitative detection of pathogenic microorganisms in food because of its rapidity and accuracy, but the PCR method cannot distinguish dead bacteria from live bacteria well, and it is also insufficient in quantitative detection. lead to inaccurate results

Images