{"doi":"10.1016/j.snb.2024.136374","title":"A single microfluidic device approach to direct isolation, purification, and amplification of cfDNA from undiluted plasma","abstract":"Circulating cell free DNA (cfDNA) is a valuable source of biomarkers for a range of medical applications including detection and monitoring of diseases. Currently, cfDNA sequence analysis must take place in a laboratory setting, due to the multiple steps required for processing including collection, purification, amplification, and analysis. Developing a point-of-care test system that combines these steps would simplify DNA processing thereby increasing diagnostic screening accessibility and enabling real-time monitoring for individual patients. Here, we have developed a system that combines multiple cfDNA processing steps into a single microfluidic-based device. This includes cfDNA collection directly from undiluted human plasma followed by purification and on chip amplification. A microelectrode array embedded within the microfluidic chip collected cfDNA through the creation of dielectrophoretic (DEP) forces followed by a wash to achieve purification. DEP utilizes differences in dielectric properties between cfDNA and plasma to preferentially induce a force on cfDNA. We then achieved on-chip amplification of collected DNA by designing a thermal cycling system to enable polymerase chain reaction (PCR) directly on the chip. This successfully consolidated the most labor-intensive steps of collection, purification, and amplification into a single device. Compared to elution of cfDNA for off-chip amplification, our on-chip PCR method improved the lower limit of detection by 3-fold and improved the total DNA yield by 5-fold. Furthermore, we demonstrate its clinical diagnostic potential by detecting KRAS mutations from a pancreatic ductal adenocarcinoma patient using only 60 µL of plasma. This paves the way for future development of a fully self-contained system facilitating the rapid detection of mutations in cfDNA. • Successful integration of DNA collection and amplification into a single device. • Electrode microarray collected cell free DNA from plasma using dielectrophoresis. • Collected DNA was PCR amplified by thermal cycling of the chip. • Pancreatic cancer-related KRAS mutations were detected in the amplified DNA. • This simplifies the most labor intensive steps for DNA analysis.","journal":"Sensors and Actuators B Chemical","year":2024,"id":445167,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":7,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9597,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1080137,"name":"Sara Evans-Dutson","orcid":"0009-0001-5517-1064","position":1,"is_corresponding":false},{"id":1261577,"name":"Jose Luis Montoya Mira","orcid":null,"position":2,"is_corresponding":false},{"id":1261078,"name":"Michael J. Heller","orcid":"0000-0001-6436-0337","position":3,"is_corresponding":false},{"id":948293,"name":"Stuart Ibsen","orcid":"0000-0001-8619-3832","position":4,"is_corresponding":false},{"id":948291,"name":"Sean Hamilton","orcid":"0000-0001-6740-290X","position":0,"is_corresponding":true}],"reference_count":23,"raw_metadata":null,"created_at":"2026-07-19T02:01:42.089235Z","pmid":"39525360","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}