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Preparation method of standard sample for establishing quantitative curve of coliform bacteria, establishment method and application of quantitative curve

A coliform, standard sample technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as errors, inconsistent sample backgrounds, affecting the accuracy of quantitative curves, etc., to achieve accurate test results and improve accuracy. sexual effect

Inactive Publication Date: 2013-03-06
INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Standard samples are used in the process of making quantitative curves. The standard samples prepared by current methods have the problem of inconsistent sample backgrounds, which affect the accuracy of quantitative curves and cause unnecessary errors when they are finally used for the detection of coliform content.

Method used

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  • Preparation method of standard sample for establishing quantitative curve of coliform bacteria, establishment method and application of quantitative curve
  • Preparation method of standard sample for establishing quantitative curve of coliform bacteria, establishment method and application of quantitative curve

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: A kind of standard sample preparation method.

[0022] 1. Solution reagent.

[0023] Activated coliform standard strain: take the Klebsiella pneumoniae standard strain (ATCC13882), inoculate it in sterilized nutrient broth, cultivate it in a constant temperature shaking incubator at 36.5±1°C for 18-24 hours, and count it for later use.

[0024] 2. Operation steps.

[0025] 1) Take melted ice cream without coliform bacteria, adjust the pH to 6.7±0.2, and make a sterile sample.

[0026] For the convenience of detection, the melted ice cream without coliform bacteria can be diluted with sterilized physiological saline according to the thickness of the sample, and then the pH is adjusted to 6.7±0.2 to make a sterile sample.

[0027] 2) Take 9 sterile samples and add a certain amount of activated coliform standard strains as shown in the table below, and adjust the pH to 6.7±0.2 to obtain 9 bacteria-rich samples.

[0028]

[0029] sample number ...

Embodiment 2

[0031] Embodiment 2: another standard sample preparation method.

[0032] 1. Solution reagent.

[0033] Activated coliform standard strain: Take the standard strain of Citrobacter freundii (ATCC43864), inoculate it in sterilized nutrient broth, cultivate it in a constant temperature shaking incubator at 36.5±1°C for 18-24 hours, and count it for later use.

[0034] 2. Operation steps.

[0035] 1) Take low-temperature yogurt without coliform bacteria, adjust the pH to 6.7±0.2, and make a sterile sample.

[0036] For the convenience of detection, according to the thickness of the sample, the low-temperature yogurt without coliform bacteria can be diluted in a certain proportion with sterilized physiological saline, and then the pH is adjusted to 6.7±0.2 to make a sterile sample.

[0037] 2) Take 7 sterile samples, add a certain amount of activated coliform standard strains as shown in the table below, and adjust the pH to 6.7±0.2 to obtain 7 bacteria-rich samples.

[0038]

...

Embodiment 3

[0041] Embodiment 3: A method for making a quantitative curve for detecting coliforms in ice cream.

[0042] 1. Instruments and equipment.

[0043] Real-time photoelectric microbial rapid detector: choose Soleris real-time photoelectric rapid microbial detector.

[0044] 2. Solution consumables.

[0045] Soleris- Coliform Medium CC-102 vials (each vial contains 5 ml of coliform selection medium and pH indicator).

[0046] 3. Operation steps.

[0047] 1) Prepare standard samples according to the standard sample preparation method described in Example 1.

[0048] 2) Inoculate 5.0 ml of the standard sample into Soleris-Coliform Medium CC-102 reagent bottle, put the reagent bottle into the Soleris real-time photoelectric microbial rapid detector for cultivation at 35±1℃, and pass the Soleris real-time photoelectric microbial The rapid detector uses colorimetry to monitor the change of the pH value in the reagent bottle. When the pH value reaches the set threshold of 5.8, recor...

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Abstract

The invention relates to a preparation method of a standard sample for establishing a quantitative curve of coliform bacteria, an establishment method and an application of the quantitative curve. Firstly a liquid sample to be detected is a sample without coliform bacteria of a same product, and the liquid sample is subject to pH value adjustment to prepare a sterile sample; a standard bacterial strain of coliform bacteria is added into the sterile sample; the pH value is adjusted to prepare a sample rich in bacteria; the sample rich in bacteria is diluted by the sterile sample for ten times in a progressive decrease tendency to prepare a standard sample. The standard sample is cultured, and detection time for the pH value reaching a certain threshold is detected. And the number of coliform bacteria in the standard sample is detected by a plate method. A quantitative curve of coliform bacteria is plotted based on the detection time and the detection result of the plate method. The liquid sample is taken and subject to pH value adjustment so as to prepare liquid to be detected; detection time is obtained by the above method; and the number of coliform bacteria in the liquid sample is obtained by comparing with the quantitative curve. The method balances the background values between samples, so the detection accuracy is improved.

Description

technical field [0001] The invention relates to a method for preparing a standard sample for making a quantitative curve, and the method for making the quantitative curve and its application are particularly suitable for the quantitative detection of coliforms in liquid samples. Background technique [0002] Conventional coliform count methods mainly include coliform Petrifilm TM Test sheet method, MPN counting method and plate counting method mentioned in the national standard GB 4789.3-2010 "National Standards of the People's Republic of China for Food Safety National Standards Food Microbiological Examination - Coliform Counting". These detection methods are time-consuming and have a large workload, which is not conducive to the rapid detection of coliform bacteria. [0003] At present, many detection methods have been developed to rapidly detect the content of coliforms by using the pH changes during the growth of coliforms. For example, the real-time photoelectric mic...

Claims

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Application Information

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IPC IPC(8): C12Q1/10C12Q1/06
Inventor 刘晓川薛志清杜丽喻东威赵源刘志楠李梅刘卫星
Owner INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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