Good evening,

I had a prior discussion with Dr. Hook that I believe was misunderstood. I would like to try and ask again. I was told my other questions were outside the scope of this forum, so I will leave those out. I hope you allow me to ask this question again.

In the previous discussion I referenced research of HPV detection in elderly women using HC2 testing (Hybrid Capture 2) and if this detection could represent non-replicable (ie non-transmissible) virus. Dr. Hook responded with the following "on occasion , most often following therapy, fragments of viral DNA may be detected by PCR type tests.  It seems unlikely however that nucleic acids detected years after clearance of infection do not represent replicative virus."

I understand that residual DNA and genetic material can exist after "clearance" or treatment, this is not my question. My question is simply this --- is it possible for at least some  HPV detections in elderly women (sexually active and non sexually active) to be a result of integrated (non-replicable non-transmissible) HPV? Can Hybrid Capture 2 pick up on integrated HPV DNA and read positive as a result with its current detection limits?   The study I am referencing used HC2 as well as external sample processing to detect HeLa cells which contain integrated hpv18 dna. I'm assuming since they are using an additional testing method to detect integrated HPV18, that HC2 is adequate to detect non-HPV18 integrated strains. Otherwise, they would test for it using other external tests as they did with Integrated HPV18.

source:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2698799/  uses HC2 and Digene Microplate Luminometer 2000 (DML 2000^™) to detect HPV

Welcome back to the Forum although I must say, it saddens me to see you perseverating on this question. I am a clinician and an epidemiologist and, as I have said before, not a molecular biologist and your questions deal with the mechanistic and procedural details of one, less and less commonly used method for detection of HPV DNA, the Hybrid Capture assay.  Please understand that both PCR and, I believe, the Hybrid Capture assays first digest the material in the specimen being tested in order to facilitate detection of HPV nucleic acids.  As I understand it, this would preclude knowing where in the cell the nucleic acids detected came from.  Integration of HPV DNA is typically accomplished by in situ hybridization.  As I tried to indicate before, your questions regarding whether or not in some patients assays for HPV are detecting non-replicative, integrative DNA are beyond the scope of this thread.

I should add , I note that you have still assiduously declined to share the reasons for your concerns.You phrase your questions as "is it possible".  Well anything is possible.  It's significance and realistic import however are unclear    I continue to wonder why you perseverate on these questions and wonder if this betrays some illogical and unwarranted focus on a hypothetical concern. 

If you have questions about the molecular biology of the Hybrid Capture assay, this is the wrong place to ask.  I suggest you ask the company.  .    EWH

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"As I understand it, this would preclude knowing where in the cell the nucleic acids detected came from..." meaning it wouldn't be possible to distinguish episomal from integrated hpv, correct? if so, I agree. - let me know if that is a correct interpretation.

I'm sorry, I thought I mentioned this in my initial correspondence with you. I am very intrigued with this because I have a (limited) background in virology and also have a personal history of previously detected high risk HPV and abnormal pap. This has since resolved, but I continue to hope that these late life detections that research keeps bringing up are either integrated HPV (no potential for progeny) or, at the very least, very low level shedding that is not clinically significant enough to result in transmission. This, I know, has yet to be proven, so I suppose I let my brain theorize.

Less and less people will be exposed to high risk HPV at younger ages due to the vaccine, meaning it is more likely to have first time exposure at a later age if still sexually active. This is especially true for men, as a large portion of them are not vaccinated, making them more susceptible to HPV at older ages from unvaccinated women. There is no data on this, but I assume first time encounters with HPV at elderly ages will be more likely to result in carcinogenic activity.

The above is my only question. Thank you for your patience, doctor.

Your interpretation is correct.

I was aware of your history.  As you point out, the issue of late intermittent shedding of HPV among persons in who the infection and any resulting PAP smear changes have resolved is the subject of ongoing research and as I've said, your questions and musings are beyond the intent or goal of this Forum.  If you wish to pursue these questions further, this is the wrong place and I would suggest you reach out to companies if you have specific questions about tests such as the Hybrid Capture or to a HPV focused molecular biologist for more general questions about the biological implications of clearance.  As I've said, prognostically, all available data suggest that persons who have cleared the virus are at little or no risk for progression of their infections or transmission to others..   I trust you'll understand- this site works to provide epidemiological and clinical information to clients.  EWH

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I understand.
I appreciate your clarifications to my prior inquiries - thank you.

Take care.