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Method for detecting sulfate reducing bacteria in water environment

A detection method and sulfate technology, applied in the measurement of color/spectral characteristics, etc., can solve the problems of limited detection sensitivity and stability, easy inactivation of biological reagents, complex testing process, etc., and achieve short detection cycle, low cost, and equipment. simple effect

Inactive Publication Date: 2012-06-20
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Other detection methods have defects to varying degrees. For example, detection through antibody-antigen binding immune reactions is easily constrained by detection conditions, and biological reagents are easily inactivated, and non-specific binding is serious.
These deficiencies limit the sensitivity and stability of the assay
Detection by biotechnology has high selectivity, but its testing process is very complicated, needs to be operated by professionals, and the cost is high

Method used

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  • Method for detecting sulfate reducing bacteria in water environment
  • Method for detecting sulfate reducing bacteria in water environment
  • Method for detecting sulfate reducing bacteria in water environment

Examples

Experimental program
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Effect test

Embodiment 1

[0015] Take the aqueous solution to be tested containing sulfate-reducing bacteria and centrifuge (centrifugal speed is 8000-10000 rpm, centrifugation time 8-10 minutes), suck out the upper layer and pour it off, take the lower layer and deposit it in the culture medium of sulfate-reducing bacteria Medium continuous culture, the culture temperature is stable at 30-35 ℃. Take it out after 3-4 days of cultivation, take the bacterial solution and centrifuge (centrifugation speed is 12000-14000 rpm, centrifugation time 10-15 minutes), and take the supernatant. Add zinc nitrate solution to the supernatant, wherein the volume ratio of supernatant to zinc nitrate is 9:1, and the concentration of zinc nitrate is 2-3mM. After reacting for 20-30 minutes, centrifuge again (centrifugal speed is 12000-14000 rpm, centrifugation time 10-15 minutes), discard the supernatant and keep the precipitated substance. Wash the precipitated substance with ultrapure water, and centrifuge again (centri...

Embodiment 2

[0020] According to the method in Example 1, detect the signal response of three kinds of bacteria (sulfate-reducing bacteria, Staphylococcus aureus, Vibrio alginolyticus) of the same concentration under the same culture time and the same ultraviolet light time conditions (see image 3 ). By measuring the absorbance of the sample at 665nm before and after ultraviolet light irradiation, the decolorization rate of different species of bacteria at the same concentration was calculated. The results showed that after deducting the background, the signal of sulfate-reducing bacteria detected by this method far exceeded that of the other two bacteria, indicating that the detection method had good selectivity. Compared with the selective detection of biological components, this method does not need to consider the activity of biomolecules and the problem of non-specific recognition.

Embodiment 3

[0022] According to the method in embodiment 1, detect a series of different concentration sulfate-reducing bacteria (from 10 1 to 10 8 cfu / mL), and the control was done without UV light irradiation. By measuring the absorbance of the sample at 665nm before and after ultraviolet irradiation, calculate the decolorization rate of sulfate-reducing bacteria in different concentrations with or without ultraviolet irradiation (see Figure 4 ). The results show that in 10 3 to 10 8 The decolorization rate increased with the concentration of sulfate-reducing bacteria in the concentration range of cfu / mL, and showed a good linear correlation. There was no significant change in the decolorization rate under the experimental conditions without light.

[0023] The invention provides a method for selective detection based on metabolites of sulfate-reducing bacteria, and the method can effectively detect the quantity and change of sulfate-reducing bacteria in a water environment. Comp...

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Abstract

The invention relates to the detection of corrosion-causing microorganisms in an environment, and particularly relates to a method for detecting sulfate reducing bacteria in a water environment. The method comprises the following steps of: after centrifuging a sample to be tested, culturing sediments in a selective culture medium for 3-4 days under anaerobic conditions; centrifuging and taking liquid supernatant; adding a zinc nitrate solution which is 1 / 9 of the volume of the liquid supernatant, uniformly mixing with the liquid supernatant, and then centrifuging to obtain sediments; adding a methylene blue solution into the sediments, uniformly mixing and then irradiating the mixed solution under an ultraviolet lamp for 1.5-2.0 hours, thus detecting the sulfate reducing bacteria in the sample to be tested through absorbance. According to the invention, a characteristic metabolic product of the sulfate reducing bacteria is combined with a detection method so as to achieve higher selectivity; and meanwhile, the defect that the culture period of the traditional method is long and the detection time is greatly shortened. According to the invention, by utilizing an optical method for detection, the method has the advantages of simple needed equipment, small operation difficulty, cheap material and very high accuracy.

Description

technical field [0001] The invention relates to the detection of corrosive microorganisms in the environment, in particular to a detection method of sulfate-reducing bacteria in the water environment. Background technique [0002] Sulfate-reducing bacteria are anaerobic microorganisms with serious corrosive hazards, which can convert sulfate ions or sulfite ions into sulfur ions through metabolism and obtain energy. The traditional detection of sulfate-reducing bacteria is through the method of maximum probability. This method needs to go through the product enrichment stage, so the required culture period is longer. Other detection methods have defects to varying degrees. For example, detection through antibody-antigen binding immune reactions is easily constrained by detection conditions, and biological reagents are easily inactivated, and non-specific binding is serious. These deficiencies limit the sensitivity and stability of the assay. Detection through biotechnolog...

Claims

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Application Information

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IPC IPC(8): G01N21/31
Inventor 戚鹏张盾万逸
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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