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Method for detecting food-derived pathogenic enterobacteria by composite fluorescence PCR technique

A composite fluorescence, technical detection technology, applied in the field of food-borne pathogenic Enterobacteriaceae, detection of pathogenic bacteria, can solve the problems of difficult biochemical identification of molecular biology detection methods, save detection cost and time, save time, Sensitive effect

Inactive Publication Date: 2008-01-30
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU +1
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  • Description
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  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows researcher to easily test different types of organism without having multiple tests at once or requiring expensive equipment such as blood smears. By measuring changes caused when certain chemicals react during an enzyme cascade called polymerase chain reactions (PC), scientists have been able to determine how well they were doing something like creating antibiotic resistance against drug resistant strains. They also found that these techniques could help predict future diseases better.

Problems solved by technology

This patented technical problem addressed by this patents relates to detecting harmful microorganisms (bacillus) in food through various techniques like polymerase chain reaction or DNA probes. However current method requires many sample preparation steps making them time-consuming and expensive compared with molecule technology. Therefore there needs an improved way to identify these organism quickly without relying on complicated chemical processes.

Method used

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  • Method for detecting food-derived pathogenic enterobacteria by composite fluorescence PCR technique
  • Method for detecting food-derived pathogenic enterobacteria by composite fluorescence PCR technique
  • Method for detecting food-derived pathogenic enterobacteria by composite fluorescence PCR technique

Examples

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Effect test

Embodiment 1

[0079] Sample: An export of fresh meat.

[0080] Use routine physiological and biochemical methods to detect suspected Shigella colonies, and then perform the following composite fluorescent PCR technology to detect foodborne pathogenic bacteria:

[0081] 1. Sample Processing

[0082] (1) Take 100 grams of the sample to be tested and pulverize it.

[0083] (2) Dissolve in 1 liter of nutrient broth and incubate at 37°C for 8 hours.

[0084] 2. DNA extraction

[0085] Take 1 mL of nutrient broth and let it stand in an ice bath for 5 minutes, then centrifuge at 12,000 rpm for 5 minutes at room temperature, discard the supernatant, add 100 μL of lysozyme solution, incubate at 37°C for 10 minutes, and add 500 μL of TE buffer , shake to mix. Add the same volume of Tris saturated phenol (pH 8.0), shake vigorously, centrifuge at 12,000 rpm for 3 minutes, take the supernatant, and repeat the phenol extraction. Take the supernatant, add 0.1 times the volume of sodium acetate (2mol / ...

Embodiment 2

[0115] Sample: Somewhere exports cookies.

[0116] Use routine physiological and biochemical methods to detect suspected colonies of Salmonella, and then perform the following composite fluorescent PCR technology to detect foodborne pathogens:

[0117] 1. Sample Processing

[0118] (1) Take 100 grams of the sample to be tested and pulverize it.

[0119] (2) Dissolve in 1 liter of nutrient broth and incubate at 37°C for 8 hours.

[0120] 2. DNA extraction

[0121] Take 1 mL of nutrient broth and let it stand in an ice bath for 5 minutes, then centrifuge at 12,000 rpm for 5 minutes at room temperature, discard the supernatant, add 100 μL of lysozyme solution, incubate at 37°C for 10 minutes, and add 500 μL of TE buffer , shake to mix. Add the same volume of Tris-saturated phenol (pH 8.0), shake vigorously, centrifuge at 12,000 rpm for 3 minutes, take the supernatant, and repeat the phenol extraction. Take the supernatant, add 0.1 times the volume of sodium acetate (2mol / L),...

Embodiment 3

[0151] Sample: Raw milk sent for inspection somewhere.

[0152] Use conventional physiological and biochemical methods to detect suspected Yersinia enterica colonies, and then perform the following composite fluorescent PCR technology to detect foodborne pathogenic bacteria:

[0153] 1. Sample Processing

[0154] (1) Take 100 grams and centrifuge.

[0155] (2) The precipitate was redissolved in 1 liter of nutrient broth and incubated at 37°C for 8 hours.

[0156] 2. DNA extraction

[0157] Take 1 mL of nutrient broth and let it stand in an ice bath for 5 minutes, then centrifuge at 12,000 rpm for 5 minutes at room temperature, discard the supernatant, add 100 μL of lysozyme solution, incubate at 37°C for 10 minutes, and add 500 μL of TE buffer , shake to mix. Add the same volume of Tris-saturated phenol (pH 8.0), shake vigorously, centrifuge at 12,000 rpm for 3 minutes, take the supernatant, and repeat the phenol extraction. Take the supernatant, add 0.1 times the volume ...

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Abstract

The invention discloses a method by using composite fluorescence PCR technique to detect food-borne pathogenic enterobacter and pertains to bacteria detection technical field. The main technic proposal is to design a primer group sequence. The pathogenic enterobacter is common pathogenic bacteria in food and imposes a serious thread to human health. The quick and accurate detection of pathogenic enterobacter in food is a main premise condition for effective prevention and control of pathogen bacteria infection. The food-borne pathogenic enterobacter required detection mainly comprises shigella, salmonella, Yersinia enterocolitica, hemorrhagic scherichia coli and scherichia coli O157: H7. The invention overcomes technical shortage in the prior art aiming at the object bacteria and provides the quick and low-cost detection method by using composite fluorescence PCR technique to detect shigella, salmonella, Yersinia enterocolitica, hemorrhagic scherichia coli and scherichia coli O157: H7. The method can use two-diode PCR reaction and primarily screen shigella, salmonella, Yersinia enterocolitica, hemorrhagic scherichia coli and scherichia coli O157: H7.

Description

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Claims

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Application Information

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Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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