ArticleBMC cancer2012
Development of a multiplex methylation-specific PCR as candidate triage test for women with an HPV-positive cervical scrape.
Article in BMC cancer, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 42 papers, 2 of them syntheses that pooled it.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
42 citing papers in PubMed, 2 syntheses or guidelines pooled it.
- The use of human papillomavirus DNA methylation in cervical intraepithelial neoplasia: A systematic review and meta-analysis.EBioMedicine · 2019Pooled it
- Detecting cervical precancer and reaching underscreened women by using HPV testing on self samples: updated meta-analyses.BMJ (Clinical research ed.) · 2018Pooled it
- Evaluation of a validated methylation triage signature for human papillomavirus positive women in the HPV FOCAL cervical cancer screening trial.International journal of cancer · 2019Trial
- Comparison of the Roche cobas® 4800 and Digene Hybrid Capture® 2 HPV tests for primary cervical cancer screening in the HPV FOCAL trial.BMC cancer · 2015Trial
- Multiplex methylation-specific PCR for distinguishing semen, saliva, and blood.International journal of legal medicine · 2026Article
- Diagnostic Accuracy of DNA Methylation Markers for Detecting Cervical Precancer and Cervical Cancer: A Systematic Review and Meta-analysis.Archives of academic emergency medicine · 2026Review
- Observational
- The diagnostic performance of a noninvasive urine-based methylation biomarkers Vimentin/POU4F2 to detect bladder carcinoma.BMC cancer · 2025Article
- Unlocking the potential of tumor-derived DNA in urine for cancer detection: methodological challenges and opportunities.Molecular oncology · 2025Review
- Bladder cancer detection in urine by novel methylation markers.Scientific reports · 2024Article
- Analytical validation and diagnostic performance of the ASCL1/ZNF582 methylation test for detection of high-grade anal intraepithelial neoplasia and anal cancer.Tumour virus research · 2024Article
- Molecular analysis for ovarian cancer detection in patient-friendly samples.Communications medicine · 2024Article
- Nanopore sequencing reveals methylation changes associated with obesity in circulating cell-free DNA from Göttingen Minipigs.Epigenetics · 2023Article
- Evaluation of DNA methylation biomarkers ASCL1 and LHX8 on HPV-positive self-collected samples from primary HPV-based screening.British journal of cancer · 2023Article
- Dynamics of methylated cell-free DNA in the urine of non-small cell lung cancer patients.Epigenetics · 2022Article
- Clinical Regression of High-Grade Cervical Intraepithelial Neoplasia Is Associated With Absence ofJournal of clinical oncology : official journal of the American Society of Clinical Oncology · 2022Article
- Technical considerations in PCR-based assay design for diagnostic DNA methylation cancer biomarkers.Clinical epigenetics · 2022Article
- Biomarker Expression in Multifocal Vulvar High-Grade Squamous Intraepithelial Lesions.Cancers · 2021Article
- DNA methylation markers have universal prognostic value for anal cancer risk in HIV-negative and HIV-positive individuals.Molecular oncology · 2021Article
- Cancer Risk Stratification of Anal Intraepithelial Neoplasia in Human Immunodeficiency Virus-Positive Men by Validated Methylation Markers Associated With Progression to Cancer.Clinical infectious diseases : an official publication of the Infectious Diseases Society of America · 2021Article
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7 authors.
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Abstract
backgroundQuantitative methylation-specific PCR (qMSP) analysis for determining the methylation status of (candidate) tumor suppressor genes has potential as objective and valuable test to triage high-risk human papillomavirus (hrHPV) positive women in cervical screening. Particularly combined methylation analysis of a panel of genes shows most promising clinical performance, with sensitivity levels that equal or exceed that of cytology. However, the wide application of such methylation marker panels is hampered by the lack of effective multiplex assays allowing simultaneous methylation detection of various targets in a single reaction. Here, we designed and analyzed a multiplex qMSP assay for three genes whose methylation was previously found to be informative for cervical (pre)cancer (i.e. CADM1, MAL and hsa-miR-124-2) as well as a reference gene β-actin. Based on our experience, we discuss the optimization of the parameters that provide a practical approach towards multiplex qMSP design.
methodsPrimers and PCR reagents were optimized for multiplex qMSP purposes and the resulting assay was analytically validated on serial dilutions of methylated DNA in unmethylated DNA, and compared with singleplex counterparts on hrHPV-positive cervical scrapings.
resultsUpon optimization, including primer redesign and primer limiting assays, the multiplex qMSP showed the same analytical performance as the singleplex qMSPs. A strong correlation between the obtained normalized ratios of the singleplex and multiplex qMSPs on cervical scrapes was found for all three markers: CADM1 (R(2) = 0.985), MAL (R(2) = 0.986) and hsa-miR-124-2 (R(2) = 0.944).
conclusionMultiplex qMSP offers a promising approach for high-throughput diagnostic analysis of the methylation status of multiple genes, which after proper design and validation can be equally specific, sensitive and reproducible as its singleplex versions.
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