Feasibility of Droplet Digital PCR Analysis of Plasma Cell-Free DNA From Kidney Transplant Patients
| العنوان: | Feasibility of Droplet Digital PCR Analysis of Plasma Cell-Free DNA From Kidney Transplant Patients |
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| المؤلفون: | David Dobnik, Maja Ravnikar, Polona Kogovšek, Barbara Jerič Kokelj, Miha Arnol, Sebastian Vencken, Matjaž Stanonik, Maja Štalekar |
| المصدر: | Frontiers in Medicine Frontiers in Medicine, Vol 8 (2021) |
| بيانات النشر: | Frontiers Media S.A., 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Oncology, medicine.medical_specialty, Medicine (General), kidney transplantation, Single-nucleotide polymorphism, DNA sequencing, droplet digital PCR, R5-920, Internal medicine, medicine, Digital polymerase chain reaction, Genotyping, Kidney transplantation, minor allele quantification, assay evaluation, business.industry, General Medicine, Brief Research Report, medicine.disease, Minor allele frequency, Transplantation, plasma cell-free DNA, graft health monitoring, Biomarker (medicine), Medicine, business |
| الوصف: | Increasing research demonstrates the potential of donor-derived cell-free DNA (dd-cfDNA) as a biomarker for monitoring the health of various solid organ transplants. Several methods have been proposed for cfDNA analysis, including real-time PCR, digital PCR, and next generation sequencing-based approaches. We sought to revise the droplet digital PCR (ddPCR)-based approach to quantify relative dd-cfDNA in plasma from kidney transplant (KTx) patients using a novel pilot set of assays targeting single nucleotide polymorphisms that have a very high potential to distinguish cfDNA from two individuals. The assays are capable of accurate quantification of down to 0.1% minor allele content when analyzing 165 ng of human DNA. We found no significant differences in the yield of extracted cfDNA using the three different commercial kits tested. More cfDNA was extracted from the plasma of KTx patients than from healthy volunteers, especially early after transplantation. The median level of donor-derived minor alleles in KTx samples was 0.35%. We found that ddPCR using the evaluated assays within specific range is suitable for analysis of KTx patients' plasma but recommend prior genotyping of donor DNA and performing reliable preamplification of cfDNA. |
| اللغة: | English |
| تدمد: | 2296-858X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4f0e845dbe598701737c929b72ee92c0 http://europepmc.org/articles/PMC8531215 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....4f0e845dbe598701737c929b72ee92c0 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 2296858X |
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