Dear All,
Is it possible to perform qPCR on Killed viral or bacterial vaccines? I read that it depends on the method of inactivation because some of the inactivation methods degrade the NA.
Does anyone have some experience doing that?
I presume that you trying to determine the Ag concentrations in inactivated vaccines.
In general Ag contents in inactivated viral vaccines are estimated by neutralization assay or particular Ag quantification by different means. In some cases the PCR dependent assay can be used to determine the mutations, for example in polio the neurovirulence proportions can be determined by MAPREC assay (https://www.who.int/biologicals/vaccines/MAPREC_SOP_Final_09112012.pdf ) again this to confirm the neurovirulence proportion not for Ag determination.
The nucleic acid based quantification may not be ideal for the quantification of vaccine content as the qPCR assay reads nucleic acids that may not differentiate infectious particle versus unpacked nucleic acids.
Dear Amro Hashish
Check these
Article Use of a Novel Real-Time PCR Assay To Detect Oral Polio Vacc...
https://www.researchgate.net/post/How_can_I_detect_of_RNA_in_inactivating_RNA_virus_vaccine https://www.researchgate.net/post/Could_possibly_use_inactive_vaccine_for_positive_control_in_PCR
Thank you all for the information.
I only need to use the inactivated bacterial vaccine as a positive control. Is that possible?
Yes it is possible based on the inactivation method as some methods destruct/fragment the nucleic acid. Just check if the vaccine is formaldehyde or binary ethyleneimine (BEI) inactivated?. Most of the huge companies nowadays are using BEI, and the NA is destructed and can´t be detected. But anyhow, if you are targeting small RNA/DNA fragment it might work in both cases.
Mahmoud M. Naguib Thank you for letting me know. I gave it a trial and it worked.
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