thanks! looks like the pcr wars are heating up - these guys make a point about being the first to report pbmc negative results
"We are the first to use an ultra-sensitive real-time PCR assay to demonstrate that occult HCV infection cannot be found in the PBMCs of patients with cryptogenic liver diseases, HCV-associated systemic vasculitis, or connective tissue diseases."
in fact, the paper by the San Francisco blood lab discussed a couple of months ago was the first to report failure to detect PBMC-HCV-RNA in serum HCV-UND negative patients. Still, overall this seems a good thing. It's hard to get a paper published on negative results alone "I carefully inspected my bathroom and found no hippopotami" unless there's a background of research showing bathrooms are chock full of hard to spot hippopotami. Pham/Castillo/Radkowski have stood pretty much unchallenged up to now. which means it should be easier to submit a paper refuting their conclusions.
However, also interesting that, like the SF paper, they did not include mitogen stimulation of the PBMCs in their protocol whereas the pioneering work of the Pham/Michalak lab has made it clear this is an important part of RNA detection in PBMCs. Unlike the SF paper, they don't even mention this issue. Nevertheless - the gauntlet has been thrown -
"we have some reservations about the existence of the so-called occult infection and we encourage publications of negative data on this topic."
A rebuttal from Spain or Newfoundland will presumably be coming along before long.
Yeah, it looks like a response (or responses) will certainly be in the offing. Though, as you mentioned, the initial response to the previous challenge was related to the lack of usage of the mitogen stimulation technique, which I imagine will be brought up again in this particular case. Then it will most likely become an issue of the validity of the mitogen technique itself.
One thing that remains relatively unchallenged so far, though, is the finding of negative-strand RNA in post-tx & post-spontaneous clearance patients hepatocytes. I wonder if any labs are actively looking in water closets to not find those hippopotamus's?
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