GB/T 43355-2023 Measurement of antiviral activity on plastics and other non-porous surfaces
WARNING - Handling and manipulation of viruses and host cells which are potentially hazardous requires a high degree of technical competence and may be subject to current national legislation and regulations. Only personnel trained in biological techniques should carry out such tests. Appropriate practices for disinfection, sterilization and personal hygiene must be strictly observed.
1 Scope
This document specifies proper methods for measuring antiviral activity on plastics and other non-porous surfaces of antiviral-treated products against specified viruses. Due to the individual sensitivities, the results of one test virus might not be applicable for other viruses.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
antiviral
state where the number of infectious virus particles on surfaces of products is reduced
3.2
antiviral agent
agent that reduces the number of infectious virus on surface of products
3.3
antiviral activity
difference in the logarithm of the infectivity titer of virus found on an antiviral-treated product and an untreated product after inoculation with and contact to virus
Note: Antiviral activity value are also called inhibitory virus activity value.
3.4
cytopathic effect
morphological change or destruction of the host cells as a result of the virus multiplication
3.5
infectivity titer of virus
number of infectious viral particles present per unit volume in a suspension
3.6
plaque
lysis formed area in a cell monolayer under semisolid medium due to infection by and multiplication of a single infectious virus
3.7
plaque forming units
PFU
unit expressed as the concentration of the infectious viral particle per unit volume (ml)
3.8
plaque assay
assay to determine the infectivity titer of virus (3.5) from PFU by using the series of dilution
4 Materials
4.1 Virus and host cell to be used for the tests
The example species of virus and host cell to be used are shown in Table 1.
Other species of virus and host cells can be used after appropriate validations, as the important virus may differ depending on the target application. If the other species are used, the name of the species and the specific reason for their use shall be included in the test report.
4.2 Reagents
Any water used shall be deionized and/or distilled and/or ultra-filtered and/or filtered with RO [reverse osmosis] and have a conductivity of <1 μS/cm.
All reagents shall be of analytical grade and/or of a grade appropriate for microbiological purposes.
4.3 Culture medium and solutions
4.3.1 General
The culture medium specified below shall be used. The medium may be obtained from commercial suppliers which shall be prepared for use in accordance with the manufacturer’s instructions.
4.3.2 Eagle's minimum essential medium (EMEM)
The composition is described in Annex A. If there are any components missing from the composition, they can be added according to Table A.1.
4.3.3 RPMI 1640 medium
The composition is described in Annex A. If there are any components missing from the composition, they can be added according to Table A.2.
4.3.4 7.5 % sodium bicarbonate solution
Select and prepare the solution using one of the two following options:
——Option 1: Prepare a 7.5 % sodium bicarbonate solution by dissolving 75 g of sodium bicarbonate in 1000 ml of water. Sterilize the solution by using a 0.22 μm membrane filter.
——Option 2: Prepare a 7.5 % sodium bicarbonate solution by sterilizing 75 g of sodium bicarbonate in a culture container with a cap closed tightly in an autoclave. Sterilize 1000 ml of water in the autoclave. Dissolve the sterilized sodium bicarbonate in the sterilized water well.
If the solution is not intended to be used immediately after preparation, then preserve it at 5 °C to 10 °C. Never use 7.5 % sodium bicarbonate solution that has been kept for longer than one month after preparation.
4.3.5 Formaldehyde solution
Prepare the formaldehyde solution by adding 100 ml of 37 % formaldehyde solution to 900 ml of water. If it is not intended to be used immediately after preparation, preserve it at 20 °C to 25 °C. Never use the formalin solution that has been kept for longer than one month after preparation.
Note: The other solution for cell fixation can be used after appropriate validation for cell fixation.
4.3.6 Methylene blue solution
Prepare the methylene blue solution by dissolving 0.375 g of the methylene blue and 62.5 μl of 1 N sodium hydroxide solutions in 1000 ml of water. If it is not intended to be used immediately after preparation, then preserve it at 20 °C to 25 °C. Never use the methylene blue solution that has been kept for longer than one month after preparation.
4.3.7 Inactivated fetal bovine serum (FBS)
Put a freezed cryopreserved fetal bovine serum in a package into a water bath at 37 °C and keep it until it defrosts. Then, raise the temperature of the water bath to 56 °C and keep it for 30 min to inactivate. Divide it into several tubes. Put them in a freezer at a temperature less than -20 °C. Just before using, put it in the water bath at 37 °C and keep it until it defrosts.
4.3.8 Growth medium
Dissolve 9.53 g of the Eagle's minimum essential medium or 10.4 g of RPMI 1640 medium and 60 mg of kanamycin sulfate in 800 ml of water. Add water to make 1000 ml of solution in total. Sterilize the solution by using a 0.22 μm membrane filter.
When L-glutamine is not included in the EMEM or RPMI 1640 medium purchased in the market, sterilizing in the autoclave may be applied. Then, before use, add L-glutamine as listed in the example of composition in Annex A.
Add 15 ml of 7.5 % sodium bicarbonate solution and 100 ml of the inactivated fetal bovine serum in the solution and mix well. If it is not intended to be used immediately after preparation, then preserve it at 5 °C to 10 °C. Never use a growth medium that has been kept for longer than one month after preparation.
4.3.9 Maintenance medium
Dissolve 9.53 g of the Eagle's minimum essential medium and 60 mg of kanamycin sulfate in 800 ml of water. Add water to make 1000 ml of solution in total. Sterilize the solution by using a 0.22 μm membrane filter.
When L-glutamine is not included in the EMEM purchased in the market, sterilizing by autoclaving may be applied. Then, before use, add L-glutamine as listed in the example of composition in Annex A.
Add 15 ml of 7.5 % sodium bicarbonate solution in the solution and mix well. If it is not intended to be used immediately after preparation, then preserve it at 5 °C to 10 °C. Never use a maintenance medium that has been kept for longer than one month after preparation.
4.3.10 Double concentration of the maintenance medium
Dissolve 19.06 g of the Eagle's minimum essential medium and 120 mg of kanamycin sulfate in 800 ml of water. Add water till there are 1000 ml of solution in total. Sterilize the solution by using a 0.22 μm membrane filter. If it is not intended to be used immediately after preparation, then preserve it at 5 °C to 10 °C. Never use a growth medium that has been kept for longer than one month after preparation. When L-glutamine is not included in the EMEM purchased on market, sterilizing by autoclaving could be applied. Then, before use, add L-glutamine as listed in the example of composition in Annex A.
4.3.11 Phosphate buffered saline [PBS(-)]
Prepare PBS(-) by dissolving 8.0 g of sodium chloride, 0.2 g of potassium chloride, 2.9 g of phosphoric acid hydrogen 2 sodium 12 hydrate and 0.2 g of phosphoric acid 2 hydrogen potassium in 1000 ml of water. Sterilize by autoclaving. If it is not intended to be used immediately after preparation, preserve it at 5 °C to 10 °C. Never use a PBS(-) that has been kept for longer than one month after preparation.
Standard
GB/T 43355-2023 Measurement of antiviral activity on plastics and other non-porous surfaces (English Version)
Standard No.
GB/T 43355-2023
Status
valid
Language
English
File Format
PDF
Word Count
10500 words
Price(USD)
315.0
Implemented on
2024-6-1
Delivery
via email in 1~3 business day
Detail of GB/T 43355-2023
Standard No.
GB/T 43355-2023
English Name
Measurement of antiviral activity on plastics and other non-porous surfaces
GB/T 43355-2023 Measurement of antiviral activity on plastics and other non-porous surfaces
WARNING - Handling and manipulation of viruses and host cells which are potentially hazardous requires a high degree of technical competence and may be subject to current national legislation and regulations. Only personnel trained in biological techniques should carry out such tests. Appropriate practices for disinfection, sterilization and personal hygiene must be strictly observed.
1 Scope
This document specifies proper methods for measuring antiviral activity on plastics and other non-porous surfaces of antiviral-treated products against specified viruses. Due to the individual sensitivities, the results of one test virus might not be applicable for other viruses.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
antiviral
state where the number of infectious virus particles on surfaces of products is reduced
3.2
antiviral agent
agent that reduces the number of infectious virus on surface of products
3.3
antiviral activity
difference in the logarithm of the infectivity titer of virus found on an antiviral-treated product and an untreated product after inoculation with and contact to virus
Note: Antiviral activity value are also called inhibitory virus activity value.
3.4
cytopathic effect
morphological change or destruction of the host cells as a result of the virus multiplication
3.5
infectivity titer of virus
number of infectious viral particles present per unit volume in a suspension
3.6
plaque
lysis formed area in a cell monolayer under semisolid medium due to infection by and multiplication of a single infectious virus
3.7
plaque forming units
PFU
unit expressed as the concentration of the infectious viral particle per unit volume (ml)
3.8
plaque assay
assay to determine the infectivity titer of virus (3.5) from PFU by using the series of dilution
4 Materials
4.1 Virus and host cell to be used for the tests
The example species of virus and host cell to be used are shown in Table 1.
Other species of virus and host cells can be used after appropriate validations, as the important virus may differ depending on the target application. If the other species are used, the name of the species and the specific reason for their use shall be included in the test report.
4.2 Reagents
Any water used shall be deionized and/or distilled and/or ultra-filtered and/or filtered with RO [reverse osmosis] and have a conductivity of <1 μS/cm.
All reagents shall be of analytical grade and/or of a grade appropriate for microbiological purposes.
4.3 Culture medium and solutions
4.3.1 General
The culture medium specified below shall be used. The medium may be obtained from commercial suppliers which shall be prepared for use in accordance with the manufacturer’s instructions.
4.3.2 Eagle's minimum essential medium (EMEM)
The composition is described in Annex A. If there are any components missing from the composition, they can be added according to Table A.1.
4.3.3 RPMI 1640 medium
The composition is described in Annex A. If there are any components missing from the composition, they can be added according to Table A.2.
4.3.4 7.5 % sodium bicarbonate solution
Select and prepare the solution using one of the two following options:
——Option 1: Prepare a 7.5 % sodium bicarbonate solution by dissolving 75 g of sodium bicarbonate in 1000 ml of water. Sterilize the solution by using a 0.22 μm membrane filter.
——Option 2: Prepare a 7.5 % sodium bicarbonate solution by sterilizing 75 g of sodium bicarbonate in a culture container with a cap closed tightly in an autoclave. Sterilize 1000 ml of water in the autoclave. Dissolve the sterilized sodium bicarbonate in the sterilized water well.
If the solution is not intended to be used immediately after preparation, then preserve it at 5 °C to 10 °C. Never use 7.5 % sodium bicarbonate solution that has been kept for longer than one month after preparation.
4.3.5 Formaldehyde solution
Prepare the formaldehyde solution by adding 100 ml of 37 % formaldehyde solution to 900 ml of water. If it is not intended to be used immediately after preparation, preserve it at 20 °C to 25 °C. Never use the formalin solution that has been kept for longer than one month after preparation.
Note: The other solution for cell fixation can be used after appropriate validation for cell fixation.
4.3.6 Methylene blue solution
Prepare the methylene blue solution by dissolving 0.375 g of the methylene blue and 62.5 μl of 1 N sodium hydroxide solutions in 1000 ml of water. If it is not intended to be used immediately after preparation, then preserve it at 20 °C to 25 °C. Never use the methylene blue solution that has been kept for longer than one month after preparation.
4.3.7 Inactivated fetal bovine serum (FBS)
Put a freezed cryopreserved fetal bovine serum in a package into a water bath at 37 °C and keep it until it defrosts. Then, raise the temperature of the water bath to 56 °C and keep it for 30 min to inactivate. Divide it into several tubes. Put them in a freezer at a temperature less than -20 °C. Just before using, put it in the water bath at 37 °C and keep it until it defrosts.
4.3.8 Growth medium
Dissolve 9.53 g of the Eagle's minimum essential medium or 10.4 g of RPMI 1640 medium and 60 mg of kanamycin sulfate in 800 ml of water. Add water to make 1000 ml of solution in total. Sterilize the solution by using a 0.22 μm membrane filter.
When L-glutamine is not included in the EMEM or RPMI 1640 medium purchased in the market, sterilizing in the autoclave may be applied. Then, before use, add L-glutamine as listed in the example of composition in Annex A.
Add 15 ml of 7.5 % sodium bicarbonate solution and 100 ml of the inactivated fetal bovine serum in the solution and mix well. If it is not intended to be used immediately after preparation, then preserve it at 5 °C to 10 °C. Never use a growth medium that has been kept for longer than one month after preparation.
4.3.9 Maintenance medium
Dissolve 9.53 g of the Eagle's minimum essential medium and 60 mg of kanamycin sulfate in 800 ml of water. Add water to make 1000 ml of solution in total. Sterilize the solution by using a 0.22 μm membrane filter.
When L-glutamine is not included in the EMEM purchased in the market, sterilizing by autoclaving may be applied. Then, before use, add L-glutamine as listed in the example of composition in Annex A.
Add 15 ml of 7.5 % sodium bicarbonate solution in the solution and mix well. If it is not intended to be used immediately after preparation, then preserve it at 5 °C to 10 °C. Never use a maintenance medium that has been kept for longer than one month after preparation.
4.3.10 Double concentration of the maintenance medium
Dissolve 19.06 g of the Eagle's minimum essential medium and 120 mg of kanamycin sulfate in 800 ml of water. Add water till there are 1000 ml of solution in total. Sterilize the solution by using a 0.22 μm membrane filter. If it is not intended to be used immediately after preparation, then preserve it at 5 °C to 10 °C. Never use a growth medium that has been kept for longer than one month after preparation. When L-glutamine is not included in the EMEM purchased on market, sterilizing by autoclaving could be applied. Then, before use, add L-glutamine as listed in the example of composition in Annex A.
4.3.11 Phosphate buffered saline [PBS(-)]
Prepare PBS(-) by dissolving 8.0 g of sodium chloride, 0.2 g of potassium chloride, 2.9 g of phosphoric acid hydrogen 2 sodium 12 hydrate and 0.2 g of phosphoric acid 2 hydrogen potassium in 1000 ml of water. Sterilize by autoclaving. If it is not intended to be used immediately after preparation, preserve it at 5 °C to 10 °C. Never use a PBS(-) that has been kept for longer than one month after preparation.
