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Chemicals—Bacterial DNA damage or repair test

Basic Information

Standard ID: GB/T 27821-2011

Standard Name:Chemicals—Bacterial DNA damage or repair test

Chinese Name: 化学品 细菌DNA损伤或修复试验方法

Standard category:National Standard (GB)

state:in force

Date of Release2011-12-30

Date of Implementation:2012-08-01

standard classification number

Standard ICS number:13.300;11.100

Standard Classification Number:Comprehensive>>Marking, packaging, transportation, storage>>A80 Marking, packaging, transportation, storage Comprehensive

associated standards

Procurement status:USEPA OPPTS 870.5500:1998 MOD

Publication information

publishing house:China Standards Press

Publication date:2012-08-01

other information

Release date:2011-12-30

drafter:Liu Qingjun, Li Chaolin, Lin Zheng, Shi Xiaoyi, Wu Weiai

Drafting unit:Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention

Focal point unit:National Technical Committee on Hazardous Chemicals Management Standardization (SAC/TC 251)

Proposing unit:National Technical Committee on Hazardous Chemicals Management Standardization (SAC/TC 251)

Publishing department:General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China Standardization Administration of China

competent authority:National Technical Committee on Hazardous Chemicals Management Standardization (SAC/TC 251)

Introduction to standards:

GB/T 27821-2011 Test Method for Bacterial DNA Damage or Repair in Chemicals GB/T27821-2011 |tt||Standard compression package decompression password: www.bzxz.net
This standard specifies the test principle, test method, test steps, test data and report for the test of bacterial DNA damage or repair in chemicals. This standard is applicable to the test of bacterial DNA damage or repair in chemicals.
This standard was drafted in accordance with the rules given in GB/T1.1-2009.
This standard is consistent with the technical content of OPPTS 870.5500:1998 Health effect test guidelines—Bacterial DNA damage or repair tests
(English version) of the Office of Prevention, Pesticides and Toxic Substances (OPPTS) of the United States Environmental Protection Agency (USEPA).
This standard has been modified as follows:
———The introduction has been changed to introduction;
———A chapter on scope has been added (see Chapter 1).
This standard was proposed and managed by the National Technical Committee for Standardization of Hazardous Chemicals Management (SAC/TC251). This standard
was drafted by: Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention.
The main drafters of this standard.

Some standard content:

ICS13.300;11.100
National Standard of the People's Republic of China
GB/T27821—2011
Chemicals
Test method for bacterial DNA damage or repair test
Chemicals-Bacterial DNA damage or repair testIssued on 2011-12-30
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China Administration of Standardization of the People's Republic of China
Implementation on 2012-08-01
This standard was drafted according to the rules given in GB/T1.12009. GB/T 27821—2011
This standard is consistent with OPPTS870.5500:1998 Health effect test guidelines - Bacterial DNA damage or repair test of the Office of Prevention, Pesticides and Toxic Substances (OPPTS) of the United States Environmental Protection Agency (USEPA) repairtcsts) (English version> The technical content is consistent, and the following structural and editorial changes have been made to this standard: the introduction part is changed to foreword
A scope chapter is added (see Chapter 1).
This standard is proposed and coordinated by the National Technical Committee for Standardization of Hazardous Chemicals Management (SAC/TC251). The drafting unit of this standard is the Institute of Occupational Health and Poisoning Control, Chinese Center for Disease Control and Prevention. The main drafters of this standard: Liu Qingjun, Li Chaolin, Lin Zheng, Chong Xiaoyi, Xi WeiI
TTTKANTKACA
GB/T 27821—2011
This standard refers to USEPA OPFTS 870.5500199B Bacterial DNA (deoxyribonucleic acid) damage or repair test (English version). This guide is one of a series of test guides developed by the Office of Prevention, Pesticides and Toxic Substances (OPPTS) of the United States Environmental Protection Agency (USEPA) to evaluate the safety of pesticides and other toxic substances. OPPTS is the result of integrating the test guidelines and requirements of two related departments. The test guidelines of these three related departments are based on the United States Code of Federal Regulations (Title 4o), the Office of Pollution Prevention and Toxic Substances (the Office of Pollution Prevention and Toxic Substances in Part R of Chapter 1) and the Office of Pollution Prevention and Toxic Substances (the Office of Pollution Prevention and Toxic Substances in Part R of Chapter 1). The testing guidelines and requirements of the Office of Pesticide Progtams (OPPT) published by the National Information Service (NTIS), and the testing guidelines published by the Organization for Economic Coaperation and Development (OECD). Control Act, 15 U.S.C. 2601) and the Federal Insecticide, Fungicide and Rodenticide Act (7 U.S.C. 136, et Seq.). To meet the requirements for test data, USEPA integrated multiple sets of guidelines into a final OPPTS test guide to reduce the differences caused by different test procedures.
KAANYKAcA
1 Scope
Test method for bacterial DNA damage or repair of chemicals GB/T 27821—2011
This standard specifies the test principle, test method, test steps, test data and report of bacterial DNA damage or repair test of chemicals.
This standard is applicable to bacterial DNA damage or repair test of chemicals. 2 Test principle
Bacterial DNA damage or repair test can be used to test DNA damage. In a set of tests of bacteria with sound repair function and repair defective strains, the repair defective strain The cell death and growth inhibition are different. This test itself is not used to detect mutation events, but to detect the genetic toxicity of the test substance by detecting the interaction between the test substance and the genetic material. By detecting the different growth inhibition effects of bacteria with sound repair function and repair-deficient strains, the test chemicals have different cell death effects on wild-type strains with sound repair function and mutant strains with repair-deficient strains. Mutant strains with repair-deficient strains are usually strains with defects in one or more regulatory DNA damage repair enzymes.
This test method can be used to detect whether a chemical can react with cell DNA to cause cell growth inhibition or lethality. The interaction between chemicals and DNA can be identified by a specific cell damage repair system. This method selects a pair of bacteria with intact and defective specific DNA damage repair genes. Since the DNA repair-deficient strain cannot repair the DNA damage caused by a certain chemical, it manifests as a tendency of cells to growth inhibition or death.
The reference substances of this method include but are not limited to chloramphenicol and methyl methanesulfonate. 3 Test methods
3.1 Method selection
Different methods can be used for the test. This method is suitable for the following two methods: a) Test on a solid matrix (diffusion test); b) Test on a liquid matrix (suspension test). 3.2 Strain selection
3.2.1 Commonly used strains
Escherichia coti potA, W3110/p3478 and Bacillus subtilis rec, H17) M45 are suitable. Other strains can also be selected.
3.2.2 Preparation and preservation of strains
The preparation and preservation of strain preservation solutions, growth conditions, identification of strains, and verification of the required phenotypes should all be based on good microbiological test methods and should be recorded.
TTTKAONTKACA
GB/T 27B21—2011
3.3 Cultivation of Bacteria
Good microbiological methods should be used to culture bacteria with fresh bacterial cultures. Record the growth phase and cell density of the cells to meet the requirements of the experimental design.
3.4 ​​Metabolic Activation
Parallel control groups with and without metabolic activation systems should be set up. The metabolic activation system used is the liver and plasma microsomal enzyme system of rodents treated with enzyme inducers (substances). Other species, tissues or methods can also be used. 3.5 Control Groups
3.5.1 Parallel Controls
Positive controls, negative controls and stimulant controls should be set up for each test. 3.5.2 Negative Controls
Negative controls should not show obvious growth inhibition (i.e., the effects on the two strains are the same). Chloramphenicol is often selected. 3,5.3 Genotype-specific controls
Methyl methanesulfonate can be used as a genotype-specific positive control for Escherichia coli tests, and Mitomyces solani can be used as a genotype-specific positive control for Bacillus subtilis tests.
3.5.4 Positive controls are used to ensure the effectiveness of the metabolic activation system. The resistance control reference material of the test, including the metabolic activation system components, is determined according to the type of activation system selected for the test. 3.5.5 Other positive controls
Other positive control reference materials can be selected. 3.6 Test substances
3.6.1 Solvents
The test substances, positive controls and negative controls should be dissolved in appropriate solvents and diluted with solvents before the test. 3.6.2. Toxic concentration
Initial tests should select a wider range of toxic concentrations, and the selection of the upper limit of the toxic concentration should be determined based on the results of cytotoxicity and the transient nature of the test substance. The cytotoxicity of the test substance affects the metabolic activation system. For non-toxic chemicals with high cytolysis, the highest dose should be determined step by step based on the actual situation. Because the results of the diffusion test are expressed as the diameter or range of the growth inhibition zone obtained, the amount of the test substance added to the culture medium or culture plate should be exactly the same. If there is a positive reaction, repeat the confirmation with a narrower concentration range. 4 Test steps
4.1 Diffusion test
4.1.1 Culture plate diffusion test
There are two methods for culture plate diffusion test: a) Add a single strain to agar for plating or smear the strain on the agar surface, and add the test substance to the filter paper on the agar surface; b) Strains with complete DNA repair function and strains with defective repair function are streaked on the agar surface, and the culture containing the test substance is placed on the agar surface to contact with the bacteria.
4.1.2 Culture plate diffusion test
In the culture plate diffusion test, the bacteria can be added to the agar for plating or smeared on the agar surface, and the test substance solution is added to the culture plate containing agar.
4.2 Suspension test
4.2.1 The bacterial suspension is inoculated with the test substance and the effect of the test substance is determined to be time-dependent or concentration-dependent based on the number of bacteria surviving (number of colonies formed).
4.2.2 For clear bacterial suspensions, a series of dilutions of the test substance may be inoculated to obtain the lowest inhibitory concentration for each strain and this may be verified by observing the appearance of visible growth after incubation. 4.2.3 Paired bacterial suspensions (usually initially isolated) may be treated with a single dose of the test substance and the different inhibitory effects may be determined by comparing the change in the rate of increase in density to obtain a positive result. 4.3 Number of cultures
If a culture diffusion test is used, each dilution should be tested in at least two separate cultures. For a suspension test, at least two independent assays should be used to determine the number of surviving cells. 4.4: Culture conditions
All cultures tested should be cultured under the same conditions, usually at 37°C for 1h-24h. 5 Test data and reporting
5.1 Results processing
5.1.1 Diffusion test
The results should be expressed as the diameter of the growth inhibition zone (in mm) or the range of the growth inhibition zone (in μm). If there are dose-response data, the same units should be used.
5.1.2 Liquid suspension test
5.1.2.1 The survival data of the strains should be given in the form of dose-response. It is advisable to give the survival percentage or survival ratio of each strain, or the relative survival ratio of two strains.
5.1.2.2 The results can also be expressed as the concentration of a certain survival rate (e.g. Dsn represents the concentration of 37% survival). The data are derived from the survival curve. The concentration should be expressed as the amount per unit volume, that is, the rate concentration. 5.1.2.3 The results can also be expressed as the minimum inhibitory concentration or the minimum lethal concentration. The minimum inhibitory concentration is obtained based on whether there is growth in liquid culture medium: the minimum lethal concentration is obtained based on the dilution of the test substance on the centrifugal culture medium. 5.1.3 Raw data
For all tests, the concentration should be expressed as the highest concentration in the test treatment. The raw data should be provided, including the actual test number, such as the net value. For diffusion tests, the diameter of the culture medium or culture plate should be stated, and the measurement results should indicate whether the diameter of the culture medium or culture plate is subtracted. In addition, it should be stated whether the test substance has a rapid, diffuse and bidirectional growth inhibition zone. If a bidirectional growth inhibition zone appears, it should be stated whether both the inner and outer zones are measured. 5.1.4 Active count
Expressed as the actual count of the culture medium at a certain dilution and the volume of the plate or the original titer (number of cells per milliliter). The final results (such as ratios, differences, survival ratios) should not be provided without providing the original test data. 5.2 Statistical evaluation
Appropriate statistical methods should be used for evaluation. 5.3 Judgment of results
5.3.1 There are multiple criteria for judging positive results. One is that the repair-deficient strain shows a statistically significant dose-related inhibition trend or mortality difference. Another criterion is that at least one test point shows a repeatable and statistically significant positive reaction. 5.3.2 If a test substance does not show a statistically significant dose-related inhibition trend or mortality difference in the repair-deficient strain, and does not show a repeatable and statistically significant positive reaction at all test points, it can be considered that the test substance has no interaction with the genetic material in the test system used.
5.3.3 Biological significance and statistical significance should be considered simultaneously in the evaluation. 5.4 Test evaluation
Bacterial DNA damage or repair tests are neither used to measure DNA damage itself nor to detect mutations. This test is to detect cell death or growth inhibition manifested by DNA damage. If the result of the DNA damage test of the test substance is positive in one test system but negative in another test system, it is difficult to judge the result without better test data. 5.5 Test Report
The test report should contain the following information:
\--The species of bacteria used;
-The growth period of the test strain:
The composition of the culture medium:
Details of the metabolic activation system used for preparation and other methods used in the test;-Treatment method, including the dosage used and the principle of dosage selection, the selection principle of positive and negative controls;-Method for determining the degree of cell death,
-Dose-response relationship (if available). 4 | | tt | ol screening chenical mutasgens; and phloxinea mutagenic red dye detected. Mutation Research. 1972,16:165-174[3] Leifer,Z. et al, An evaluation ol bacterial DNA repair lests for predicting genotaxicity and carcinogcnicity,A report of the U, S. EPA's Gene-Tox Program. Mutation Research, 198l,87:2ll-297[4J Slater, EE, et al, Rapid detection of mutagcns and carcinogens. Cancer Rcscarch. 171,3l:970-9732 Plate Diffusion Test
In the plate diffusion test, the bacteria are either added to the agar plate or smeared on the agar surface, and the test substance solution is added to the agar-containing plate.
4.2 Suspension Test
4.2.1 The bacterial suspension is inoculated with the test substance, and the effect of the test substance is judged to be time-dependent or concentration-dependent based on the number of bacterial viable (colony formation).
4.2.2 For clear bacterial suspensions, a series of test substance dilutions may be inoculated to obtain the lowest inhibitory concentration for each strain, which can be verified by observing whether visible growth occurs after incubation. 4.2.3 Paired bacterial suspensions (usually initially isolated) may be treated with a single dose of the test substance, and the different inhibitory effects may be judged by comparing the changes in the rate of increase in density, resulting in a positive result. 4.3 Number of Cultures
If the culture diffusion test is selected, each dilution should be tested in at least two separate cultures. For in vivo suspension tests, at least two independent assays should be used to determine the number of surviving cells. 4.4: Culture conditions wwW.bzxz.Net
All cultures in the test should be cultured under the same conditions, usually at 37°C for 1 h-24 h. 5 Test data and reporting
5.1 Handling of results
5.1.1 Diffusion tests
The results should be expressed as the diameter of the growth inhibition zone (in mm) or the extent of the growth inhibition zone (in μm). If dose-response data are available, the same units should be used.
5.1.2 Liquid suspension tests
5.1.2.1 The survival data of the strains should be presented in a dose-response format. It is advisable to present the percentage survival or survival ratio of each strain, or the relative survival ratio of two strains.
5.1.2.2 The results may also be presented as the concentration that gives a certain survival rate (e.g. Dsn represents the concentration that gives 37% survival). The data are derived from the survival curve. The concentration should be expressed as the amount per unit volume, i.e., the concentration per unit volume. 5.1.2.3 The results can also be expressed as the minimum inhibitory concentration or the minimum lethal concentration. The minimum inhibitory concentration is obtained based on whether there is viable growth in the liquid culture medium; the minimum lethal concentration is obtained based on the dilution concentration of the test substance in the semi-peripheral culture medium. 5.1.3 Raw data
For all tests, the concentration should be expressed as the highest concentration in the test treatment. The raw data should be provided, including the actual number of tests, such as the net value. For diffusion tests, the diameter of the culture medium or culture plate should be stated, and the measurement results should indicate whether the diameter of the culture medium or culture plate was subtracted. In addition, it should be stated whether the test substance has a rapid, diffuse and bidirectional growth inhibition zone. If a bidirectional growth inhibition zone is present, it should be stated whether both the inner and outer zones have been measured. 5.1.4 Active counts
Expressed as the actual counts of cultured blood at a certain dilution and the volume of the plate or the original titer (number of cells per milliliter). The final results (such as ratios, differences, survival ratios) should not be provided without providing the original test data. 5.2 Statistical evaluation
Appropriate statistical methods should be selected for evaluation. 5.3 Judgment of results
5.3.1 There are multiple criteria for judging positive results. One is that the repair-deficient strain shows a statistically significant dose-related inhibition trend or death difference. Another criterion is that at least one test point shows a repeatable and statistically significant positive reaction. 5.3.2 If a test substance does not show a statistically significant dose-related inhibition trend or death difference on the repair-deficient strain, and does not show a repeatable and statistically significant positive reaction at all test points, it can be considered that the test substance has no interaction with the genetic material in the test system used.
5.3.3 Biological significance and statistical significance should be considered simultaneously in the evaluation. 5.4 Test Evaluation
Bacterial DNA damage or repair tests are not designed to measure DNA damage itself, nor are they designed to detect mutations. The test is to detect cell death or growth inhibition as a result of DNA damage. If the DNA damage test result of the test substance is positive in one test system, but negative in another test system, it is difficult to interpret the result without better test data. 5.5 Test Report
The test report should contain the following information:
- The species of bacteria used;
- The growth period of the test strain;
The composition of the culture medium;
Details of the metabolic activation system used to prepare and other methods used in the test;
Treatment method, including the dose used and the rationale for dose selection, positive and negative controls;
- Method for determining the degree of cell death,
- Dose-response relationship (if available). 4 | | tt | ol screening chenical mutasgens; and phloxinea mutagenic red dye detected. Mutation Research. 1972,16:165-174[3] Leifer,Z. et al, An evaluation ol bacterial DNA repair lests for predicting genotaxicity and carcinogcnicity,A report of the U, S. EPA's Gene-Tox Program. Mutation Research, 198l,87:2ll-297[4J Slater, EE, et al, Rapid detection of mutagcns and carcinogens. Cancer Rcscarch. 171,3l:970-9732 Plate Diffusion Test
In the plate diffusion test, the bacteria are either added to the agar plate or smeared on the agar surface, and the test substance solution is added to the agar-containing plate.
4.2 Suspension Test
4.2.1 The bacterial suspension is inoculated with the test substance, and the effect of the test substance is judged to be time-dependent or concentration-dependent based on the number of bacterial viable (colony formation).
4.2.2 For clear bacterial suspensions, a series of test substance dilutions may be inoculated to obtain the lowest inhibitory concentration for each strain, which can be verified by observing whether visible growth occurs after incubation. 4.2.3 Paired bacterial suspensions (usually initially isolated) may be treated with a single dose of the test substance, and the different inhibitory effects may be judged by comparing the changes in the rate of increase in density, resulting in a positive result. 4.3 Number of Cultures
If the culture diffusion test is selected, each dilution should be tested in at least two separate cultures. For in vivo suspension tests, at least two independent assays should be used to determine the number of surviving cells. 4.4: Culture conditions
All cultures in the test should be cultured under the same conditions, usually at 37°C for 1 h-24 h. 5 Test data and reporting
5.1 Handling of results
5.1.1 Diffusion tests
The results should be expressed as the diameter of the growth inhibition zone (in mm) or the extent of the growth inhibition zone (in μm). If dose-response data are available, the same units should be used.
5.1.2 Liquid suspension tests
5.1.2.1 The survival data of the strains should be presented in a dose-response format. It is advisable to present the percentage survival or survival ratio of each strain, or the relative survival ratio of two strains.
5.1.2.2 The results may also be presented as the concentration that gives a certain survival rate (e.g. Dsn represents the concentration that gives 37% survival). The data are derived from the survival curve. The concentration should be expressed as the amount per unit volume, i.e., the concentration per unit volume. 5.1.2.3 The results can also be expressed as the minimum inhibitory concentration or the minimum lethal concentration. The minimum inhibitory concentration is obtained based on whether there is viable growth in the liquid culture medium; the minimum lethal concentration is obtained based on the dilution concentration of the test substance in the semi-peripheral culture medium. 5.1.3 Raw data
For all tests, the concentration should be expressed as the highest concentration in the test treatment. The raw data should be provided, including the actual number of tests, such as the net value. For diffusion tests, the diameter of the culture medium or culture plate should be stated, and the measurement results should indicate whether the diameter of the culture medium or culture plate was subtracted. In addition, it should be stated whether the test substance has a rapid, diffuse and bidirectional growth inhibition zone. If a bidirectional growth inhibition zone is present, it should be stated whether both the inner and outer zones have been measured. 5.1.4 Active counts
Expressed as the actual counts of cultured blood at a certain dilution and the volume of the plate or the original titer (number of cells per milliliter). The final results (such as ratios, differences, survival ratios) should not be provided without providing the original test data. 5.2 Statistical evaluation
Appropriate statistical methods should be selected for evaluation. 5.3 Judgment of results
5.3.1 There are multiple criteria for judging positive results. One is that the repair-deficient strain shows a statistically significant dose-related inhibition trend or death difference. Another criterion is that at least one test point shows a repeatable and statistically significant positive reaction. 5.3.2 If a test substance does not show a statistically significant dose-related inhibition trend or death difference on the repair-deficient strain, and does not show a repeatable and statistically significant positive reaction at all test points, it can be considered that the test substance has no interaction with the genetic material in the test system used.
5.3.3 Biological significance and statistical significance should be considered simultaneously in the evaluation. 5.4 Test Evaluation
Bacterial DNA damage or repair tests are not designed to measure DNA damage itself, nor are they designed to detect mutations. The test is to detect cell death or growth inhibition as a result of DNA damage. If the DNA damage test result of the test substance is positive in one test system, but negative in another test system, it is difficult to interpret the result without better test data. 5.5 Test Report
The test report should contain the following information:
- The species of bacteria used;
- The growth period of the test strain;
The composition of the culture medium;
Details of the metabolic activation system used to prepare and other methods used in the test;
Treatment method, including the dose used and the rationale for dose selection, positive and negative controls;
- Method for determining the degree of cell death,
- Dose-response relationship (if available). 4 | | tt | ol screening chenical mutasgens; and phloxinea mutagenic red dye detected. Mutation Research. 1972,16:165-174[3] Leifer,Z. et al, An evaluation ol bacterial DNA repair lests for predicting genotaxicity and carcinogcnicity,A report of the U, S. EPA's Gene-Tox Program. Mutation Research, 198l,87:2ll-297[4J Slater, EE, et al, Rapid detection of mutagcns and carcinogens. Cancer Rcscarch. 171,3l:970-9733 Raw data
For all tests, the concentration should be expressed as the highest concentration in the test treatment. Raw data should be provided, including the actual test number, such as the net value. For diffusion tests, the diameter of the culture medium or culture plate should be stated, and the measurement result should indicate whether the diameter of the culture medium or culture plate was subtracted. In addition, it should be stated whether the test substance shows rapid, diffuse and bidirectional growth inhibition zones. If bidirectional growth inhibition zones appear, it should be stated whether both the inner and outer zones were measured. 5.1.4 Viability count
It should be expressed as the actual count of the culture medium at a certain dilution and the volume of the plate or the original titer (number of cells per milliliter). The final result (such as ratio, difference, survival ratio) should not be provided without providing the raw test data. 5.2 Statistical evaluation
Appropriate statistical methods should be used for evaluation. 5.3 Judgment of results
5.3.1 There are multiple criteria for judging positive results. One is that the repair-deficient strain shows a statistically significant dose-related inhibition trend or death difference. Another criterion is that a repeatable and statistically significant positive reaction appears at least at one test point. 5.3.2 If a test substance does not show a statistically significant dose-related inhibition trend or death difference on the repair-deficient strain, and does not show a repeatable and statistically significant positive reaction at all test points, it can be considered that the test substance has no interaction with the genetic material in the test system used.
5.3.3 Biological significance and statistical significance should be considered simultaneously in the evaluation. 5.4 Test evaluation
Bacterial DNA damage or repair tests are neither used to measure DNA damage itself nor to detect mutations. This test detects cell death or growth inhibition manifested by DNA damage. If the result of the DNA damage test of the test substance is positive in a certain test system, but negative in another test system, the result is difficult to judge without better test data. 5.5 Test Report
The test report should contain the following information:
\--The species of bacteria used;
-The growth period of the test strain;
The composition of the culture medium;
Detailed information on the metabolic activation system used to prepare the test and other methods used in the test;-Treatment method, including the dosage used and the principle of dosage selection, the principle of selection of positive and negative controls;-Method for determining the degree of cell death,
-Dose-response relationship (if available). 4 | | tt | ol screening chenical mutasgens; and phloxinea mutagenic red dye detected. Mutation Research. 1972,16:165-174[3] Leifer,Z. et al, An evaluation ol bacterial DNA repair lests for predicting genotaxicity and carcinogcnicity,A report of the U, S. EPA's Gene-Tox Program. Mutation Research, 198l,87:2ll-297[4J Slater, EE, et al, Rapid detection of mutagcns and carcinogens. Cancer Rcscarch. 171,3l:970-9733 Raw data
For all tests, the concentration should be expressed as the highest concentration in the test treatment. Raw data should be provided, including the actual test number, such as the net value. For diffusion tests, the diameter of the culture medium or culture plate should be stated, and the measurement result should indicate whether the diameter of the culture medium or culture plate was subtracted. In addition, it should be stated whether the test substance shows rapid, diffuse and bidirectional growth inhibition zones. If bidirectional growth inhibition zones appear, it should be stated whether both the inner and outer zones were measured. 5.1.4 Viability count
It should be expressed as the actual count of the culture medium at a certain dilution and the volume of the plate or the original titer (number of cells per milliliter). The final result (such as ratio, difference, survival ratio) should not be provided without providing the raw test data. 5.2 Statistical evaluation
Appropriate statistical methods should be used for evaluation. 5.3 Judgment of results
5.3.1 There are multiple criteria for judging positive results. One is that the repair-deficient strain shows a statistically significant dose-related inhibition trend or death difference. Another criterion is that a repeatable and statistically significant positive reaction appears at least at one test point. 5.3.2 If a test substance does not show a statistically significant dose-related inhibition trend or death difference on the repair-deficient strain, and does not show a repeatable and statistically significant positive reaction at all test points, it can be considered that the test substance has no interaction with the genetic material in the test system used.
5.3.3 Biological significance and statistical significance should be considered simultaneously in the evaluation. 5.4 Test evaluation
Bacterial DNA damage or repair tests are neither used to measure DNA damage itself nor to detect mutations. This test detects cell death or growth inhibition manifested by DNA damage. If the result of the DNA damage test of the test substance is positive in a certain test system, but negative in another test system, the result is difficult to judge without better test data. 5.5 Test Report
The test report should contain the following information:
\--The species of bacteria used;
-The growth period of the test strain;
The composition of the culture medium;
Detailed information on the metabolic activation system used to prepare the test and other methods used in the test;-Treatment method, including the dosage used and the principle of dosage selection, the principle of selection of positive and negative controls;-Method for determining the degree of cell death,
-Dose-response relationship (if available). 4 | | tt | ol screening chenical mutasgens; and phloxinea mutagenic red dye detected. Mutation Research. 1972,16:165-174[3] Leifer,Z. et al, An evaluation ol bacterial DNA repair lests for predicting genotaxicity and carcinogcnicity,A report of the U, S. EPA's Gene-Tox Program. Mutation Research, 198l,87:2ll-297[4J Slater, EE, et al, Rapid detection of mutagcns and carcinogens. Cancer Rcscarch. 171,3l:970-973E, et al, Rapid detectiun of mutagcns and carcinogens. Cancer Rcscarch. 171,3l:970-973E, et al, Rapid detectiun of mutagcns and carcinogens. Cancer Rcscarch. 171,3l:970-973
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