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Method for rapidly detecting coliforms by means of spectrophotometry and application of method

A spectrophotometry and coliform technology, applied in the field of microbial detection, can solve the problems of high cost of molecular biology detection method, low temperature preservation of paper medium, unsuitable for field operation, etc. Reduced, easy-to-use effects

Active Publication Date: 2014-07-23
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, coliform detection technologies in the prior art, such as filter membrane method, enzyme substrate method, and molecular biology detection method, have the defects of high cost and cumbersome operation; in addition, the current paper method is also commonly used in most fields. method, but the paper medium has the disadvantage of needing low temperature storage, so it is not suitable for field work

Method used

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  • Method for rapidly detecting coliforms by means of spectrophotometry and application of method
  • Method for rapidly detecting coliforms by means of spectrophotometry and application of method
  • Method for rapidly detecting coliforms by means of spectrophotometry and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A method for rapid detection of coliform bacteria by spectrophotometry, the steps are as follows:

[0044] (1) Solution preparation:

[0045] Lactose fermentation test medium: 50μL 2% (mass concentration) bromocresol purple solution, 4g tryptone, 0.5g lactose, 0.5gD-mannitol, 0.75gNaCl, 0.02g sodium lauryl sulfate, 0.01g sodium deoxycholate, 100mL distilled water, pH 6.8, sterilized at 115°C for 15 minutes before use;

[0046] TTC solution: Add sterile water to TTC to make a 3% (mass concentration) solution, keep it away from light and wait for use.

[0047] (2) Detection process:

[0048] ① Preparation of homogeneous sample solution:

[0049]Aseptic operation Take 25g or 25mL of the sample to be tested, put it into a triangular flask containing 225mL of sterile saline (with a number of glass beads inside), homogenize at 8000r / min~10000r / min for 1min~2min, and make the volume ratio 1:10 Homogenized sample solution (if the detection limit of the liquid sample to be t...

Embodiment 2

[0055] A method for rapid detection of coliform bacteria by spectrophotometry, the steps are as follows:

[0056] (1) Solution preparation:

[0057] Lactose fermentation test medium: 90μL 2% (mass concentration) bromocresol purple solution, 4g tryptone, 0.5g lactose, 0.5gD-mannitol, 0.75gNaCl, 0.02g sodium lauryl sulfate, 0.01g sodium deoxycholate, 100mL distilled water, pH7.2, 115°C, sterilized for 15min before use;

[0058] TTC solution: Add sterile water to TTC to make a 1.5% (mass concentration) solution, keep it away from light and wait for use.

[0059] (2) Detection process:

[0060] ① Preparation of homogeneous sample solution:

[0061] Aseptic operation Take 25g or 25mL of the sample to be tested, put it into a triangular flask containing 225mL of sterile saline (with a number of glass beads inside), homogenize at 8000r / min~10000r / min for 1min~2min, and make the volume ratio 1:10 Homogenized sample solution (if the detection limit of the liquid sample to be tested...

Embodiment 3

[0067] A method for rapid detection of coliform bacteria by spectrophotometry, the steps are as follows:

[0068] (1) Solution preparation:

[0069] Lactose fermentation test medium: 60μL 2% (mass concentration) bromocresol purple solution, 4g tryptone, 0.5g lactose, 0.5gD-mannitol, 0.75gNaCl, 0.02g sodium lauryl sulfate, 0.01g sodium deoxycholate, 100mL distilled water, pH6.9, 115℃, sterilized for 15min before use;

[0070] TTC solution: Add sterile water to TTC to make a 2% (mass concentration) solution, keep it away from light and wait for use.

[0071] (3) Detection process:

[0072] ① Preparation of homogeneous sample solution:

[0073] Aseptic operation Take 25g or 25mL of the sample to be tested, put it into a triangular flask containing 225mL of sterile saline (with a number of glass beads inside), homogenize at 8000r / min~10000r / min for 1min~2min, and make the volume ratio 1:10 Homogenized sample solution (if the detection limit of the liquid sample to be tested is...

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Abstract

The invention relates to a method for rapidly detecting coliforms by means of spectrophotometry. By utilizing the characteristic that coliforms can be utilized to ferment lactose at 37 DEG C to produce acid and gas and taking a self-made culture medium as the main role, bromcresol purple serving as an indicator and triphenyltetrazoliumchloride (TTC) solutions are added, after coliforms is fermented and cultured, red precipitates can be produced and culture liquid turns from purple into yellow; the characteristic that the change degree of the color of the fermentation liquid is in positive correlation with the quantity of coliforms contained in the solutions is utilized, the content of coliforms in sample liquid to be detected can be obtained. Compared with an existing national standard method, the method has the advantages that the detection time is greatly shortened, the quantitative value of coliforms in the sample can be detected when coliforms are cultured for 60-400 minutes after sample adding, the trial range is wide, the requirement on short detection time required by various detection fields is met, and the method has good application prospect in the field of rapid detection in the future.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection and relates to the detection of coliforms, in particular to a method and application for rapid detection of coliforms by spectrophotometry. Background technique [0002] Coliform bacteria refer to Gram-negative non-bacillus bacteria that can ferment lactose, produce acid and gas, aerobic and facultative anaerobic at 37°C, mainly derived from human and animal feces. The content of coliform bacteria in a sample is an important indicator for testing food hygiene and safety. At present, the commonly used detection methods include traditional fermentation method, rapid disk method, filter membrane method, enzyme substrate method, polymerase chain method, fluorescence in situ hybridization method, but the detection time of these known detection methods still needs 24h-72h. etc. For production testing of industrial wastewater, food, etc., the detection cycle of coliform bacteria is long,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 黄琳张朝正郭刚程雅文刘晓博刘爽王炼
Owner TIANJIN UNIV OF SCI & TECH
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