Since several references indicate the presence of HCV in blood cells and that RNA extraction from whole blood samples leads to higher amounts of viral RNA. So why is plasma/serum routinely used instead of whole blood?
Is this related to RNA extraction methods? Would the use of total RNA extraction methods form whole blood samples result in too much RNA that would complicate PCR optimization?
Thank you
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You are right Salma. HCV and many other viruses do persist in blood cells. However for diagnostics serum or plasma is preferred since they contain less cell, thus less host DNA/RNA is obtained after nucleic extraction. The lesser the host nucleic acids, the better will be the detection. Furthermore, virus circulating in the serum or plasma are believed to have originated from liver (at least for HBV). If you are interested in studying viruses from blood cell, you have to isolate them from enriched blood cells, from which serum/plasma is properly washed away. You may get different strains/genotypes of virus in the serum/plasma and blood cells from the same individuals. This is called 'compartmentalization' and a lot of papers on HCV compartmentalization could be found in pubmed search.
Best wishes,
SD
Thank you very much Dr Datta for your answer.
Assuming that the an extraction method capable of specifically extracting viral RNA from whole blood was used- as opposed to total RNA extraction- Would that potentially increase the amount of RNA going into PCR and according limits of detection?
if you are capable of specifically extracting viral RNA from blood cells then you can have better qPCR results, I hope.
SD
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