Rapid detection method of environmental microorganisms

Abstract

The invention discloses a rapid detection method of environmental microorganisms. The method comprises the following steps: (1) collecting the microorganisms, namely collecting the microorganisms by an air filter net; (2) extracting microbial genomes; (3) carrying out 16SrDNA amplification on bacteria, and carrying out 18SrDNA amplification on fungi; (4) sequencing ribosome deoxyribonucleic acid (rDNA); (5) analyzing data comprising a decontamination sequence, a low-quality sequence, a merged high-similarity sequence and a sequence annotation; and (6) building an environmental bacteria database and an environmental fungi database, wherein the environmental bacteria database comprises category information, sequence information and abundance information, and the environmental fungi database comprises category information, sequence information and abundance information. The rapid detection method is especially suitable for general investigation of the environmental microorganisms in a clean area, and is faster, more accurate and more sensitive than a conventional culture-based detection technology; the detection cost and time are greatly reduced; the capacity of monitoring and controlling microorganism pollution is significantly improved. On the other hand, a microbial molecular level mark is also provided by gene sequencing data, and thus the rapid detection method has important value for reviewing and judging a microorganism source and accurately identifying a pollution source.

Description

technical field [0001] The invention relates to a detection method for environmental microorganisms, which can improve the detection range of microorganisms, save detection costs and greatly shorten detection time. Background technique [0002] Microorganisms are the most widely distributed and the largest class of organisms in nature. They are small in size, fast in reproduction and strong in adaptability. Some microorganisms invade the human body through the digestive tract, such as Salmonella, Vibrio parahaemolyticus, Listeria monocytogenes, etc. They can cause gastrointestinal diseases, and in severe cases, they can cause damage to the liver, brain, kidneys, and even death. The danger of mold lies in the toxins it produces, such as aflatoxins, fumonisins, etc., which are highly toxic and carcinogenic. In addition, if microorganisms and pyrogens directly invade the human body through mucous membranes or blood vessels, they will cause more serious and direct harm to hum...

AI Technical Summary

Problems solved by technology

[0005] However, the microbial testing capabilities of most food and drug manufacturers in my country still remain at the cell level

These inspection methods are established to adapt to product release inspection, and there is generally insufficient capacity in the inspection of microbial contamination process control

At the same time, the traditional microbiological method can only cultivate no more than 10% of the microorganisms in the environment, not only the detection time is long, the accuracy of the results is poor, but also the efficiency is low and the workload is heavy

Although the industry has developed some fully automated microbial identification instruments, their identification range is limited by known microbial databases

Therefore, traditional methods cannot truly reveal the diversity of microbial communities

Generally speaking, the food and drug industry is in a relatively passive position regarding microbial contamination, not only lacking the means to investigate microbial infection patterns (such as microbial species, quantity and pathways), but also lacking quantitative risk assessment indicators

Blindly adopting excessive disinfection (sterilization) measures in actual production is not only blindly inefficient, but also increases environmental pollution and personnel safety risks

For products exceeding the standard, there is no other way but to be scrapped and destroyed, and it is difficult to play a guiding role in subsequent production

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