ArticleCellular oncology (Dordrecht, Netherlands)2017
Digital droplet PCR (ddPCR) for the detection and quantification of HPV 16, 18, 33 and 45 - a short report.
Article in Cellular oncology (Dordrecht, Netherlands), 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers, 1 of them a synthesis that pooled it.
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Who cites it
35 citing papers in PubMed, 1 synthesis or guideline pooled it, 64 citations in OpenAlex.
- The prevalence of human papillomavirus in pediatric tonsils: a systematic review of the literature.Journal of otolaryngology - head & neck surgery = Le Journal d'oto-rhino-laryngologie et de chirurgie cervico-faciale · 2018Pooled it
- Type-specific detection and quantification of circulating tumor HPV DNA by digital PCR in surgically resected cervical cancer.Journal of gynecologic oncology · 2026Article
- Detection of papillomavirus DNA in uterine flushing samples from healthy mares reveals diverse genotypes with differing host ranges.Scientific reports · 2026Article
- A droplet digital PCR assay targeting 16 human papillomavirus genotypes.Access microbiology · 2026Article
- Can Oncogenic Animal Viruses Pose a Threat to Humans?Pathogens (Basel, Switzerland) · 2025Review
- Accurate identification of bovine deltapapillomavirus in equine sarcoids by ddPCR.Scientific reports · 2025Article
- Advances in human papillomavirus detection for cervical cancer screening and diagnosis: challenges of conventional methods and opportunities for emergent tools.Analytical methods : advancing methods and applications · 2025Review
- Operational Feasibility and Performance of mRPA Versus PCR for the Detection of HPV Types 16, 18, and 52 in Clinical Samples: An Exploratory Study.International journal of women's health · 2025Article
- Evolving HPV diagnostics: current practice and future frontiers.Frontiers in cellular and infection microbiology · 2025Review
- Molecular findings and virological assessment of bladder papillomavirus infection in cattle.The veterinary quarterly · 2024Article
- Detection and quantification by molecular techniques of early infection by Lawsonia intracellularis in suckling piglets.Porcine health management · 2024Article
- The Role of Biomarkers in HPV-Positive Head and Neck Squamous Cell Carcinoma: Towards Precision Medicine.Diagnostics (Basel, Switzerland) · 2024Review
- Applying improved ddPCR to reliable quantification of MPXV in clinical settings.Microbiology spectrum · 2024Article
- Portable, and ultrasensitive HR-HPV tests based on nucleic acid biosensors.Frontiers in cellular and infection microbiology · 2024Review
- Droplet digital PCR as alternative to microbiological culture forFrontiers in cellular and infection microbiology · 2024Article
- Development of a sensitive droplet digital PCR according to the HPV infection specificity in Chinese population.BMC cancer · 2023Article
- Development of a reverse-transcription droplet digital PCR method for quantitative detection ofHeliyon · 2023Article
- Droplet digital PCR application for the detection of SARS-CoV-2 in air sample.Frontiers in public health · 2023Article
- False Negative Results in Cervical Cancer Screening-Risks, Reasons and Implications for Clinical Practice and Public Health.Diagnostics (Basel, Switzerland) · 2022Review
- Detection of Circulating HPV16 DNA as a Biomarker for Cervical Cancer by a Bead-Based HPV Genotyping Assay.Microbiology spectrum · 2022Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
purposeHuman papilloma virus (HPV) infection is associated with several anogenital malignancies. Here, we set out to evaluate digital droplet PCR (ddPCR) as a tool for HPV 16, 18, 33 and 45 viral load quantification and, in addition, to compare the efficacy of the ddPCR assay for HPV 16 detection with that of quantitative real-time PCR (qPCR).
methodsClinical samples, positive for HPV genotypes 16, 18, 33 and 45 were analyzed for viral load using ddPCR. Sample DNA was cleaved before droplet generation and PCR. Droplets positive for VIC and FAM fluorescence were read in a QX200 Droplet reader™ (BIO-RAD) after which the viral load was calculated using Quantasoft software.
resultsWe found that DNAs extracted from formalin fixed paraffin embedded (FFPE) tissue samples yielded lower amplification signals compared to those obtained from liquid based cytology (LBC) samples, but they were clearly distinguishable from negative background signals. The viral limit of detection was 1.6 copies of HPV 16, 2.8 copies of HPV 18, 4.6 copies of HPV 33 and 1.6 copies of HPV 45. The mean inter-assay coefficients of variability (CV) for the assays ranged from 3.4 to 7.0%, and the mean intra-assay CV from 2.6 to 8.2%. The viral load in the different cohorts of tumor samples ranged from 154 to 340,200 copies for HPV 16, 244 to 31,300 copies for HPV 18 and 738 to 69,100 copies for HPV 33. One sample positive for HPV 45 contained 1331 viral copies. When comparing qPCR data with ddPCR copy number data, the qPCR values were found to be 1 to 31 times higher.
conclusionsSeparation of fragments in nanodroplets may facilitate the amplification of fragmented human and viral DNA. The method of digital droplet PCR may, thus, provide a new and promising tool for evaluating the HPV viral load in clinical samples.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.