Anti-interference rapid detection and reagent for microbe

A detection method and microbial technology, applied in the direction of chemiluminescence/bioluminescence, analysis of materials through chemical reactions, bacteria, etc., can solve the problems of interference from preservatives or residual disinfectants, insufficient sensitivity, etc., to shorten the detection time, Effect of Improving Detection Sensitivity

Inactive Publication Date: 2005-10-12
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY +1
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  • Claims
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Problems solved by technology

[0007] The purpose of the present invention is to overcome the problem of insufficient sensitivity of the current ATP bioluminescence method, solve the interference problem of some preservatives or residual disinfe

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  • Anti-interference rapid detection and reagent for microbe

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Embodiment 1

[0058] Embodiment 1: Bacteria liquid large sample rapid determination in bottled drinking natural mineral water

[0059] Because bottled drinking natural mineral water is usually disinfected with ozone, an anti-ozone bacterial liquid large-sample detection medium is selected:

[0060] a. Preparation of culture medium: prepare 50mL / bottle of three-material liquid culture medium according to the instruction of anti-ozone bacteria liquid large sample detection medium, divide into 250mL triangular flasks, prepare other supporting supplies, and then sterilize;

[0061] b. Cultivation of microorganisms: In a 100-level ultra-clean workbench, add 100mL of the sample to be tested into a 250mL Erlenmeyer flask filled with cooled 50mL / bottle of three-material medium, shake and mix, and then culture at 35-37°C 6h;

[0062] c. Microbial cell ATP extraction method: take 10mL of culture medium, centrifuge at 10000r / min for 5 minutes, remove the supernatant, add 1mL of microbial cell ATP rel...

example 2

[0068] Example 2: Rapid Determination of MPN Method for Yeast and Mold in Juice Drinks

[0069] Juice drinks are usually added with preservatives to prolong the shelf life, but the freshly produced products contain very low bacteria, so to determine the microbial count of samples with low bacterial counts containing preservatives, the MPN method can be used. ), to test the sample:

[0070] a. Preparation of culture medium: according to the national standard coliform number multi-tube fermentation method (9-tube method), prepare anti-corrosion agent-type fungal liquid large sample detection medium (no need to add a small inverted tube to the test tube), 10mL per tube, and prepare Other supporting supplies, and then sterilized;

[0071] b. Cultivation of microorganisms: In a 100-class ultra-clean workbench, add the mixed and dispersed samples into the measurement system according to the multi-tube fermentation method for the number of coliforms in the national standard, shake a...

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Abstract

A fast detection method of microbe anti- interference includes using large sample method or MPN method to enrich bacteria for microbe culture media sample, processing prepared sample with cell ATP release agent Ec and adding luciferase - luciferin reagent and deinhibitor for ATP luminous detection, confirming that sample is bacteria carrier positive if sample luminous pulse counts CPMs is greater than control sample solution CPMck, checking MPN table to obtain bacteria content in sample. The method can raise detection sensitivity by 1000 times comparing to normal method.

Description

[technical field] [0001] The invention belongs to a method and a reagent for detecting and measuring microorganisms, in particular to a method for rapidly detecting microorganisms in a sample containing less bacteria by using bioluminescence and a reagent used in the method. [Background technique] [0002] At present, the detection of the number of bacteria, mold and yeast, which is the most common routine detection item for microorganisms, still uses the agar plate counting method established by Pasteur more than 100 years ago. Traditional microbial detection methods rely on special microbial culture media to isolate and culture the living microorganisms in samples. These methods are sensitive and intuitive, and can provide information on the number and types of microorganisms in food at the same time. However, the conventional method takes 1 to 5 days from the beginning of cultivation to the colonies visible to the naked eye, and the operation is cumbersome, limited to the...

Claims

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Application Information

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IPC IPC(8): C12N1/20G01N21/76
Inventor 吴清平张菊梅吴慧清郭伟鹏
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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