Happy Monday ResearchGate community,
I need standard DNA samples for 14 high-risk HPV genotypes to calibrate an in-house multiplex real-time PCR assay.
I have contacted NIBSC and the International HPV reference center without any response. I am actively following them up, but wonder is there any other path you would suggest me to follow.
Eagerly look forward to your kind suggestions.
Best wishes,
Negar
Hello Negar Fakhari
Of the 14 high-risk HPV subtypes, two of them can be easily procured. If you have access to a cell culture facility then look for HeLa, SiHa and CaSki cell lines. These cervical cancer cell lines have copies of HPV DNA in them. HeLa is positive for HPV-18 and both SiHa and CaSki are positive for HPV-16.
Getting the positive control or standard for other high-risk subtypes is a little challenging. In our case what we did was, we screened through all available samples with consensus HPV primers and then sequenced them to identify the possible presence of other high-risk genotypes. If you get any positives there then they can be cloned and used for further analysis.
Hope this helps.
Best!
Hi Dr. Sinduvadi Ramesh,
Thank you for your response.
After consulting with several impactful scientists in the HPV diagnostics niche, I learned that the NIBSC is an internationally-approved source that can provide quantified standard samples with a background of human cells for the genotypes included in the vaccine.
Another institute that might be helpful is the HPV international reference center. They can provide a limited number of clones for various types.
Many thanks again Pushkal Sinduvadi Ramesh
Best wishes,
Negar
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