Academic Journal

Quantification of human papillomavirus cell-free DNA from low-volume blood plasma samples by digital PCR

التفاصيل البيبلوغرافية
العنوان: Quantification of human papillomavirus cell-free DNA from low-volume blood plasma samples by digital PCR
المؤلفون: Fabian Rosing, Matthias Meier, Lea Schroeder, Simon Laban, Thomas Hoffmann, Andreas Kaufmann, Oliver Siefer, Nora Wuerdemann, Jens Peter Klußmann, Thorsten Rieckmann, Yvonne Alt, Daniel L. Faden, Tim Waterboer, Daniela Höfler
المصدر: Microbiology Spectrum, Vol 12, Iss 7 (2024)
بيانات النشر: American Society for Microbiology, 2024.
سنة النشر: 2024
المجموعة: LCC:Microbiology
مصطلحات موضوعية: HPV, liquid biopsy, digital PCR, OPC, cfDNA, early detection, Microbiology, QR1-502
الوصف: ABSTRACT The incidence rate of human papillomavirus-driven oropharyngeal cancer (HPV-OPC) is increasing in countries with high human development index. HPV cell-free DNA (cfDNA) isolated from 3 to 4 mL blood plasma has been successfully used for therapy surveillance. A highly discussed application of HPV-cfDNA is early detection of HPV-OPC. This requires sensitive and specific cfDNA detection as cfDNA levels can be very low. To study the predictive power of pre-diagnostic HPV-cfDNA, archived samples from epidemiological cohorts with limited plasma volume are an important source. To establish a cfDNA detection workflow for low plasma volumes, we compared cfDNA purification methods [MagNA Pure 96 (MP96) and QIAamp ccfDNA/RNA] and digital PCR systems (Biorad QX200 and QIAGEN QIAcuity One). Final assay validation included 65 low-volume plasma samples from oropharyngeal cancer (OPC) patients with defined HPV status stored for 2–9 years. MP96 yielded a 28% higher cfDNA isolation efficiency in comparison to QIAamp. Both digital PCR systems showed comparable analytical sensitivity (6–17 copies for HPV16 and HPV33), but QIAcuity detected both types in the same assay. In the validation set, the assay had 80% sensitivity (n = 28/35) for HPV16 and HPV33 and a specificity of 97% (n = 29/30). In samples with ≥750 µL plasma, the sensitivity was 85% (n = 17/20), while in samples with
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 2165-0497
Relation: https://doaj.org/toc/2165-0497
DOI: 10.1128/spectrum.00024-24
URL الوصول: https://doaj.org/article/48533d1238d7461a9b378bb4cdffd83c
رقم الانضمام: edsdoj.48533d1238d7461a9b378bb4cdffd83c
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:21650497
DOI:10.1128/spectrum.00024-24