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Simple method for detecting water genotoxicity substances in source water

A genotoxic, water source water technology, applied in biochemical equipment and methods, microbial determination/inspection, color/spectral property measurement, etc. Low sterility requirements and time-saving effect

Active Publication Date: 2011-09-14
GUANGDONG INST OF MICROORGANISM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Based on the above materials, it can be seen that the Ames test and the SOS / Umu test method are all methods for detecting the damage degree of DNA, and the Ames mutagenic test is a widely used test method at present, but due to the large number of strains required for it, it is difficult to operate Complicated, high detection costs limit its application; the standard method of SOS / Umu test is very cumbersome to operate, and it needs to transform the culture plate multiple times during the operation, and the conditions for the configuration and preservation of the culture medium required by the bacteria are relatively harsh. Reagents, the specificity of biomarkers is not significant enough and the cost is quite high

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  • Simple method for detecting water genotoxicity substances in source water
  • Simple method for detecting water genotoxicity substances in source water
  • Simple method for detecting water genotoxicity substances in source water

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 A Simple Method for Detecting Genotoxic Substances in Source Water

[0026] The simple method for detecting genotoxic substances in source water described in this embodiment mainly includes the following steps:

[0027] 1) Inoculate bacterial strain TA1535 (purchased from DSMZ, German Culture Collection Center, deposit number is DSM 9274) into liquid LB medium for overnight culture (14-18 hours) to obtain a bacterial suspension, wherein the formula of LB medium is: peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L.

[0028] 2) Adjust the concentration of the bacterial suspension to make it OD 595 Reach 0.28-0.30.

[0029] Pipette 100 μl of overnight cultured TA1535 bacterial suspension into a 1.5ml centrifuge tube, centrifuge at 5000rpm for 5min to collect the bacteria, dilute to 1.0ml with sterile water, and measure its OD 595 value. Then use bacteria and sterile water to adjust its concentration to make its OD 595Reach 0.28-0.30. Take 600μl of t...

Embodiment 2

[0042] Embodiment 2: Simple method of the present invention and SOS / umu standard method reliability, sensitivity difference comparison

[0043] In order to compare the reliability and sensitivity of the two methods, two different methods were used to determine the original DNA damage substances in tap water in a certain place in Guangzhou under the same test conditions, and the test was repeated three times.

[0044] The simple method used in this example is the same as that in Example 1, and the comparison results with the SOS / umu standard method are specifically shown in Table 1.

[0045] Table 1 Comparison of two methods under the same test conditions

[0046]

[0047] It can be seen from Table 1 that there is no significant difference in the IR values ​​of the two methods at the level of α=0.05.

Embodiment 3

[0048] Embodiment 3: Simple method of the present invention compares with SOS / umu standard method sensitivity difference

[0049] In order to compare the reliability and sensitivity of the two methods, two different methods were used to determine the original DNA damage substances in tap water in a certain place in Guangzhou under the same test conditions, and the test was repeated three times.

[0050] The simple method for detecting genotoxic substances in source water described in this embodiment has the following steps:

[0051] 1) Inoculate the bacterial strain TA1535 (purchased from the German Culture Collection Center DSMZ, the deposit number is DSM 9274) into the nutrient broth medium for overnight culture (14-16 hours) to obtain a bacterial suspension, wherein the formula of the nutrient broth medium For: peptone 10g, beef extract 3g, sodium chloride 5g, distilled water 1000mL, pH7.4.

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Abstract

The present invention discloses a simple method for detecting water genotoxicity substances in source water, comprising the following steps of: inoculating strain TA1535 into liquid LB or culture medium of nutrient broth, carrying out shaking cultivation, adjusting the concentration of bacterial suspension with sterile water to lead OD595 to be 0.2 to 0.3, and obtaining bacterial suspension; taking water sample concentrates with at least four concentration gradient, respectively adding the water sample concentrates into the bacterial suspension, carrying out shaking cultivation, adopting DMSO as solvent control, and adopting 4-NQO as positive control; adding B-Buffer after shaking cultivation, mixing evenly, adding chloroform, shaking and mixing evenly; adding ONPG solution, incubating and developing for 30 to 50 minutes; adding Na2CO3 solution, stopping developing, measuring the OD420 and OD550 of the solution after developing; calculating enzymatic activity of samples and solvent control, and calculating inductivity IR. The simple method for detecting water genotoxicity substances in source water can save many steps and large amount of time under the premise of keeping routinization of test operation and having accurate and sensitive test result.

Description

technical field [0001] The invention relates to the technical field of environmental toxicology, in particular to a simple method for detecting genotoxic substances in source water. Background technique [0002] my country's current drinking water quality standards and hygienic norms are basically in line with the three major foreign standards. In particular, the revised health norms and urban water supply standards of the Ministry of Health and the Ministry of Construction have greatly increased the toxicological indicators of organic pollutants. However, there are no relevant laws and regulations on the implementation and implementation of drinking water standards in my country, and it is difficult to implement. In addition, due to biological genotoxicity, especially the detection of medium and long-term biological genotoxicity requires relatively high technical difficulty, long time and high cost. Therefore, my country usually only monitors conventional sensory, microbia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02G01N21/31
Inventor 孙国萍张国霞郭俊曾国驱许玫英张家强
Owner GUANGDONG INST OF MICROORGANISM
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