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Coliform single-tube quantitative rapid detection method

A technology of coliform group and detection method, applied in the detection field, can solve the problems of a large number of inoculation test tubes, a large error, a long detection time, etc., and achieve the effects of high sensitivity, rapid detection results, and shortening of the incubation time.

Inactive Publication Date: 2009-12-30
JIMEI UNIV
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Problems solved by technology

There are two main defects in this method: the first defect is that the detection time is long, and the method needs to be incubated for 18-24 hours after the sample is inoculated; the second defect is that the maximum possible number method is used for quantification, resulting in a large number of inoculated test tubes , generally need to inoculate 9 or 15 test tubes, and the error of the maximum probable number method is relatively large

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  • Coliform single-tube quantitative rapid detection method

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example 1

[0025] Example 1: Prepare ONPG culture solution, divide it into test tubes, 10ml per test tube, inoculate 1ml of sample into the culture solution, incubate at 42°C for 10 hours, measure the absorbance value of the culture solution at 405nm at 1.20 with a spectrophotometer after centrifugation , according to the corresponding formula of absorbance and bacterial concentration under this condition, the number of coliform bacteria in this sample is 4.1×10 4 cfu / ml.

example 2

[0026] Example 2: Prepare ONPG culture solution, divide it into test tubes, 10ml per test tube, inoculate 1ml of sample into the culture solution, cultivate at 42°C for 12 hours, measure the absorbance value of the culture solution at 405nm with a spectrophotometer after centrifugation is 1.10 , according to the corresponding formula of absorbance and bacterial concentration under this condition, the number of coliform bacteria in this sample is 186cfu / ml.

example 3

[0027] Example 3: Prepare ONPG culture solution, divide it into test tubes, 10ml per test tube, inoculate 1ml of sample into the culture solution, incubate at 42°C for 14 hours, measure the absorbance value of the culture solution at 405nm with a spectrophotometer after centrifugation is 1.30 , according to the corresponding formula of absorbance and bacterial concentration under this condition, the number of coliform bacteria in this sample is 16cfu / ml.

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Abstract

The invention discloses a coliform single-tube quantitative rapid detection method which comprises the steps: preparing specific detection culture medium; inoculating sample; culturing the inoculated sample for 8-15h at the temperature of 35-44DEG C; centrifugating for 5-10min at the speed of 8000-12000rpm; measuring absorbance value at the part of 405nm of supernate by a spectrophotometer; and calculating bacteria concentration according to formula. The method utilizes the spectrophotometer to measure the absorbance value of culture solution, and is more sensitive than visual evaluation, thus greatly shortening the culturing time. By adopting the single-tube quantitative detecting technique, one simple needs a tube of culture solution, so that a great deal of reagent and consumable materials can be saved compared with most probable number method; furthermore, the detection method is more rapid and stabler, and has high sensitivity.

Description

technical field [0001] The invention relates to a detection method, in particular to a single-tube quantitative rapid detection method for coliform bacteria. Background technique [0002] At present, the most probable number method (MPN) is used for the detection of coliform bacteria in samples based on color production technology at home and abroad, that is, single-tube qualitative and multi-tube quantitative, and the detection methods are visual inspection of color. There are two main defects in this method: the first defect is that the detection time is long, and the method needs to be incubated for 18-24 hours after the sample is inoculated; the second defect is that the maximum possible number method is used for quantification, resulting in a large number of inoculated test tubes , generally need to inoculate 9 or 15 test tubes, and the error of the maximum probable number method is relatively large. Contents of the invention [0003] The purpose of the present inven...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/06G01N21/31C12R1/19
Inventor 鄢庆枇张小东庄峙厦王小如
Owner JIMEI UNIV
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