Fast quantitative detection method of shigella contained in sludge

Abstract

The invention relates to a fast quantitative detection method of shigella contained in sludge, belonging to the technical field of pathogenicbacteria detection in biotechnologies. The fast quantitative detection method of the shigella contained in the sludge comprises the following steps of: (1) carrying out gradient dilution and front enriched culture on a sludge sample; (2) carrying out selectively enriched culture; (3) carrying out DNA extraction; (4) carrying out PCR (Polymerase Chain Reaction) amplification and amplified product detection; and (5) calculating to obtain a result by utilizing MPN (Most Probable Number) counting software. The invention realizes the fast quantitative detection of the shigella contained in the sludge by adopting a method combining an MPN method with specific PCR detection and can prevent the disturbance of other microbes through selective enrichment and specific PCR, thereby enhancing the sensitivity and the specificity of detection; and in addition, the invention also has the advantages of shortening the detection time, enhancing the detection efficiency and increasing the quantity of detection samples.

Description

technical field [0001] The invention discloses a rapid quantitative detection method for Shigella in sludge environment, and belongs to the technical field of pathogenic bacteria detection in biotechnology. Background technique [0002] Shigella ( Shigella ) bacteria, commonly known as Shigella, rank first in the proportion of the original bacteria that cause infectious diarrhea in my country. They are highly infectious and serious Gram-negative enteropathogenic bacteria. Every year, 160 million people around the world suffer from bacterial infections, and about 1.1 million people die, most of whom are children under 5 years old. Since domestic sewage contains many pathogenic microorganisms, such as bacteria, viruses, protozoa, parasites, etc. from the intestinal tract, most of these microorganisms will enter the residual sludge with sewage treatment. During the agricultural land use of sludge, It diffuses into the environment through various ways such as polluting surface...

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Problems solved by technology

However, when this method is applied to the detection of sludge samples, on the one hand, due to the complex composition of sludge and a large number of microorganisms, it will interfere with the detection sensitivity; Various disadvantages

There are also other patents reporting Shigella detection methods, such as a Shigella detection primer and detection method invented by South China Agricultural University (CN101845493A) and a detection kit for Salmonella and Shigella in feed invented by Cao Jijuan and its detection method (CN101624624A), but none of them can meet the requirements of rapid quantitative detection of Shigella in sludge samples

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