Ecological biological recognition method based on MPN algorithm

Abstract

The invention discloses an ecological biological recognition method based on an MPN algorithm, and the method comprises the following steps: sample collection: collecting a water sample by using a pre-sterilized sampling bottle, and storing the collected water sample in an ice box; gradient dilution of the water sample: diluting by adopting three continuous gradients; enrichment culture of the water sample: respectively adding the samples with three dilution degrees into enrichment liquid, uniformly mixing, taking 5 parts of the enrichment liquid for each dilution degree, and culturing in a constant-temperature incubator at 42 DEG C for 24 hours; dNA extraction: extracting and purifying bacterial DNA by adopting a glass bead method, a magnetic bead method, a microcolumn method or an organic solvent extraction method; pCR amplification: taking the extracted DNA sample as a template, and carrying out PCR amplification by using a salmonella specific primer; and carrying out statistical analysis through an MPN algorithm to calculate the bacteria content of the water sample. According to the present invention, with the MPN method, the content of the salmonella living body in the water can be quantitatively detected, the detection specificity is improved, the interference of other microorganisms is avoided, the operation is simple, convenient and feasible, and the method is suitable for the detection of various water samples such as surface water, sewage and the like.

Description

technical field [0001] The invention relates to the technical field of biological identification, in particular to an ecological biological identification method based on the MPN algorithm. Background technique [0002] Salmonella is widely distributed, and the consensus among the academic circles is that it is the type of bacteria with the highest degree of contamination in food. In the statistics of bacterial food poisoning cases around the world, the number of cases caused by Salmonella often takes the first or second place. So far, more than 2600 Salmonella serotypes have been reported worldwide. Typhoid fever, paratyphoid fever, and fever caused by Salmonella are the most common diseases in developing countries, and even lead to death in areas with poor sanitation conditions. According to reports, about 94 million cases of gastroenteritis and 155,000 deaths worldwide are caused by Salmonella each year, 85% of which are caused by food. [0003] There are many species o...

AI Technical Summary

Problems solved by technology

[0003] There are many types of Salmonella, and their characters often vary in the water environment, and the actual biochemical reaction results often do not match the types in the Salmonella identification table, resulting in detection failures

The biggest disadvantage of the existing detection and identification methods is that they cannot distinguish between living and dead bacteria, and cannot deeply study the impact of Salmonella in the water environment on human health.

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