Hello
I am trying to find a method to inactivate swine influenza viruses that are produced in eggs, most of the items gives formalin, but I find a specific protocol, I need suggestions and ideas if someone is familiar with this method, I await your suggestions
thank you
Yes influenza virus can be inactivated at very low doses of formalin (0.02%) , you can check this publication for more details
http://jcm.asm.org/content/48/3/928.full
Yes low concentrations of formalin inactivate the virus. Most of formalin (pure and buffered) inactivate the virus
Yes influenza virus can be inactivated at very low doses of formalin (0.02%) , you can check this publication for more details
http://jcm.asm.org/content/48/3/928.full
Yes, most of viruses, including influenza virus can be inactivated with low concentration of formalin.
Thank you for all of you
I need a functional protocol to follow if any of you worked with this type of protocol
thank you
Usually, 0,1% of formalin solution, time of inactivation should be optimised (4 - 12 hours at 25C with constant mixing are usually enought).
Why formalin? I prefer beta-propiolactone (0,1% of 99% pure BPL).
Yes, formalin in low concentration inactivates the virus as suggested by others before me, has expressed.
Formalin will form methyl cross linkage in protein and fix the protein, so it is expected that it will fix the viral protein and inactivate virus.
- I have been using 0.1 to 0.3 % Formalin ( 37.5% strength) to inactivate chicken egg propagated AI viruses. In my observation we need to standardise a method for our own environment because a lot depends on yolk contamination of AF harvest. Lower concentrations are effective only for very clean harvests but higher concentration ( 0.3% , 4C, 24hr ) ensures complete inactivation of even not so clean allantoic fluids. However HA titres before and after inactivation must be checked to trend formalin Lots because some Lots of formalin reduce HA titre more than others probably owing to the quality of methanol in the formalin solution. Hope it helps.
suggest you to carry on kinetics of inactivation with 0.02-0.3 % conc. formalin and choose the ideal conc. which inactivates the virus without the loss of HA activity.
I have inactivated purified virus using the following conditions:
Prepare formalin by diluting stock formaldehyde solution (37% w/w containing 10-15% methanol as preservative) 1:40 in calcium saline (0.1ml stock formaldehyde solution plus 4.0ml calcium saline buffer).
Sterilize by passing through a 0.45µm filter.
Prepare purified virus to give 1mg/ml (~40,000 HAU) in calcium borate saline.
Mix 1 part of dilute formalin (1/40) with 99 parts of concentrated virus for a final concentration of formaldehyde 1/4000 and protein concentration < 1mg/ml.
Store at 4oC for at least 3 days.
Follow “Virus Innocuity Test” protocol to check for virus inactivation.
you can also inactivate virus with Beta-propiolactone. Can send you a protocol if you need it.
Does anyone observed precipitation (white string type material) while adding formaldehyde to clarified influenza harvest?
regarding Priyabrata question in the given answers,I view white cloudy precipitation after inactivation with Formaldehyde . I make a 0.2% Formaldehyde stock solution and take my needed volumes from that.
need to add that this precipitation does not pass through 0.22 Filter. Do You have any idea what could be the reason ?
thanks
No idea as not done and now not attached to any lab. I am suggesting some one to look into.
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