Separation, Purification, and Detection of cfDNA in a Microfluidic Device

Kye, Hyeon Gi; Ahrberg, Christian D.; Park, Byeong Seon; Lee, Jong Min; Chung, Bong Geun

doi:10.1007/s13206-020-4208-1

Cited by 14 publications

(8 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The micromixer plateaued at 2, 1, and 0.8 s at 1, 2, and 3 Hz, respectively, and mixing indices reached 96.3 ± 3.1% at 2 Hz. These high values could be due to the three larger membranes around the chamber versus those of the prior design with four small membranes (σ = 96%) and the Tesla mixer (σ = 94%); thus, a stronger vortex-like flow could be generated. Therefore, the saturation time and mixing index of this micromixer were comparable with or even superior to the micromixers in the previous studies, , in which the Tesla mixer was also developed for cfDNA isolation …”

Section: Resultsmentioning

confidence: 99%

Isolation and Quantification of Methylated Cell-Free DNA in Plasma on an Integrated Microfluidic System

et al. 2022

View full text Add to dashboard Cite

Methylated cell-free DNA (cfDNA) has been deemed a promising biomarker for ovarian cancer (OvCa) prognosis and therapy selection. However, exploring the methylation profiles of tumor suppressor genes in cfDNA remains a challenge due to their extremely low concentrations and complicated protocols, as well as methodological constraints. In this study, an integrated microfluidic system was developed to automatically (1) capture methylated cfDNA in plasma by magnetic beads coated with the methyl-CpG-binding domain and (2) quantify the methylation level of tumor suppressor genes by on-chip quantitative polymerase chain reaction (qPCR). For capturing methylated cfDNA from a very small amount of plasma, samples along with beads were mixed in a new micromixer to enhance the capture rate. With a high capture rate (72%) and a limit of quantification of 0.1 pg/μL (3 orders of magnitude lower than that of the benchtop method), the compact system could detect the methylated cfDNA from only 20 μL of plasma sample in 2 h. Furthermore, the dynamic range, from 0.1 to 2000 pg/μL of methylated cfDNA, spans the physiological range in plasma, signifying that this device has great potential for personalized medicine in OvCa.

show abstract

Section: Resultsmentioning

confidence: 99%

Isolation and Quantification of Methylated Cell-Free DNA in Plasma on an Integrated Microfluidic System

et al. 2022

View full text Add to dashboard Cite

show abstract

“…The most common microfluidic method for DNA extraction from blood or plasma is miniaturized versions of SPE with silica microbeads [171,178,179]. Different microfluidic modules have been integrated with microfluidic SPE for DNA analysis, including PCR detection [180,181], bacteria pre-concentration [182,183], plasma separation [184,185], droplet digital PCR [181], or digital microfluidics [186]. Alternatives to microfluidic SPE have been reported.…”

Section: Nucleic Acid Biomarkersmentioning

confidence: 99%

“…However, for the analysis of RNA or cfDNA, it is essential to separate the plasma from the blood cells to eliminate enzymatic inhibitors, such as heme [168] or hemoglobin [169]. Microfluidic devices for nucleic acids analysis that integrate plasma separation from blood have been reported [180,[196][197][198]. These examples clearly demonstrate a current trend and natural evolution in developing integrated solutions to separate plasma and detect nucleic acids in the same device from whole blood samples.…”

Section: Nucleic Acid Biomarkersmentioning

confidence: 99%

“…As for detecting nucleic acid biomarkers, CD platforms [196], membrane filtration [197], sedimentation [198], and hydrodynamic forces [180] have been used to separate plasma from blood. For example, a double‐rotation axes centrifugal microfluidic platform was shown to detect the DNA of hepatitis B in serum extracted from 500 µl of whole blood.…”

Section: Integrated Microfluidic Systems: Plasma Preparation and Biom...mentioning

confidence: 99%

See 1 more Smart Citation

Microfluidic systems for the analysis of blood‐derived molecular biomarkers

Chavez-Pineda¹,

Rodriguez‐Moncayo²,

Cedillo-Alcantar³

et al. 2022

Electrophoresis

View full text Add to dashboard Cite

Biomarkers are relevant indicators of the physiological state of an individual.Although biomarkers can be found in diseased tissue and different biofluids, sampling from blood plasma is relatively easy and less invasive. Among the molecular biomarkers that can be found circulating in plasma are proteins, metabolites, nucleic acids, and exosomes. Some of these plasma-circulating biomarkers are now employed for patient stratification in a broad range of diseases with high sensitivity and specificity and are useful in early diagnosis, initial risk assessment, and therapy selection. However, there is a pressing need to develop novel approaches for biomarker analysis that can be translated into clinical or other settings without complex methodologies or instrumentation. Microfluidics has been touted as a promising technology to carry out this task because it offers high-throughput, automation, multiplexed detection, and portability, possibly overcoming the bottleneck that prevent the translation of novel biomarkers to the point-of-care (POC). Here, we provide a review of the microfluidic systems that have been engineered to detect circulating molecular biomarkers in blood plasma. We also review the different microfluidic approaches for plasma enrichment, which are now being integrated with microfluidic-based biomarker analyzers. Such integration should lead to

show abstract

“…Separation of nucleic acids can provide information about the presence of genomic biomarker for disease diagnosis [16,17]. For instance, the size distribution of cell-free DNA (cfDNA) was employed to predict ICU mortality of patients [18] and long dsRNA from plant leaves was electrophoretically separated to confirm viral infection in plants [19].…”

Section: Introductionmentioning

confidence: 99%

On-chip Paper Electrophoresis for Ultrafast Screening of Infectious Diseases

Kang

et al. 2021

BioChip J

View full text Add to dashboard Cite

The outbreak of new viral strains promotes advances in universal diagnostic techniques for detecting infectious diseases with unknown viral sequence. Long double-stranded RNA (dsRNA), a hallmark of infections, serves as a virus marker for prompt detection of viruses with unknown genomes. Here, we report on-chip paper electrophoresis for ultrafast screening of infectious diseases. Negatively charged RNAs pass through the micro and nanoscale pores of cellulose in order of size under an external electric field applied to the paper microfluidic channel. Quantitative separation of long dsRNA mimicking poly I:C was analyzed from 1.67 to 33 ng·μL−1, which is close to the viral dsRNA concentration in infected cells. This paper-based capillary electrophoresis chip (paper CE chip) can provide a new diagnostic platform for ultrafast viral disease detection at the point-of-care (POC) level.

show abstract

Separation, Purification, and Detection of cfDNA in a Microfluidic Device

Cited by 14 publications

References 36 publications

Isolation and Quantification of Methylated Cell-Free DNA in Plasma on an Integrated Microfluidic System

Isolation and Quantification of Methylated Cell-Free DNA in Plasma on an Integrated Microfluidic System

Microfluidic systems for the analysis of blood‐derived molecular biomarkers

On-chip Paper Electrophoresis for Ultrafast Screening of Infectious Diseases

Contact Info

Product

Resources

About