We are trying to detect a Virus using a sandwich lateral flow assay. We are targeting the epitopes on the envelop protein of the virus. Is there anyway I can inactivate the disinfect the virus, but still retain its ability to bind to the antibodies. The reason for inactivating the virus is to handle it outside the hood.
Thank You
Greetings Gangula,
You can try inactivation by UV-B. The radiation will damage the DNA/RNA and therefore the virus won't be able to replicate, but the epitopes will remain intact.
More information here: Article Inactivation of Viruses on Surfaces by Ultraviolet Germicida...
Thank you very much Ivan for your prompt response. Are you also aware of any chemical methods which we can follow in our lab. We don't have a UV radiation facility.
Hi Gangula,
The traditional method for inactivating a virus while retaining the antigenic properties of the virion is heat inactivation. This process is as simple as it sounds - you heat the sample for a set period of time at a specific temperature. It is likely that there is published data for the time and temperature for the virus you are working with but it is important to remember the following points:
1) You must considered other pathogens that may be present (eg HIV, Hep B etc in blood). You should also investigate whether the protocol with inactivate these pathogens
2) You must ensure your sample reaches the required temperature by monitoring the temperature of a mock sample (ideally a mock tube containing similar matrix with a temperature probe in it to indicate if your samples reached the required temperature)
3) You should always try and mitigate against instances where the samples were not properly inactivated - this is usually achieved by wearing basic PPE and maintaing good laboratory practice when working with the samples downstream + disposing of samples in a suitable manner. It is also important to remember that many published protocols only achieve a set knock down (4-6 logs) rather than sterilisation (ie complete inactivation of all virions) so maintaining good practice is ALWAYS recommended.
4) Some viruses are extremely thermostable so you cannot assume that standard protocols (eg 56C-60C for 30-60 mins) will work for your virus. In these instances, you can add chemicals to mitigate the risk of incomplete inactivation.
If there is no heat inactivation data for the virus you are working with, then you'll have to generate your own by performing analysis on varying temperatures for varying times on mock samples spiked with known quantities of virus then trying to culture virus from the samples after "inactivation".
Here are a few articles that may help
Article Effects of Viral Hemorrhagic Fever Inactivation Methods on t...
Article Comparative Thermostability of West Nile, St. Louis Encephal...
Article Virus-specific thermostability and heat inactivation profile...
Hope that helps
Barry
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