A three-in-one microfluidic droplet digital PCR platform for absolute quantitative analysis of DNA

Ren, Yulin; Ji, Jingcheng; Zhang, Haoqing; Cao, Lei; Hu, J. F.; Xing, Feng; Li, Zedong

doi:10.1039/d3lc00107e

Cited by 8 publications

(4 citation statements)

References 27 publications

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“…It is feasible that a single microfluidic chip may be able to perform the entire UR-ddPCR process after UR-DNA extraction-i.e., generating droplets, thermal cycling, and reading droplets-at greatly increased speed and sample parallelization. Microfluidic chips already exist that perform some or all of these steps, albeit none designed for ultrarapid speed 29,30 . We estimate that an UR-ddPCR microfluidics chip could decrease the time required for profiling a single sample by more than half and may eliminate the time difference between profiling multiple samples in parallel versus a single sample.…”

Section: Discussionmentioning

confidence: 99%

Ultra-Rapid Droplet Digital PCR Enables Intraoperative Tumor Quantification

Murphy,

Bianchini,

Smith

et al. 2024

Preprint

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The diagnosis and treatment of tumors often depends on molecular-genetic data. However, rapid and iterative access to molecular data is not currently feasible during surgery, complicating intraoperative diagnosis and precluding measurement of tumor cell burdens at surgical margins to guide resections. To address this gap, we developed Ultra-Rapid droplet digital PCR (UR-ddPCR), which can be completed in 15 minutes from tissue to result with an accuracy comparable to standard ddPCR. We demonstrate UR-ddPCR assays for the IDH1 R132H and BRAF V600E clonal mutations that are present in many low-grade gliomas and melanomas, respectively. We illustrate the clinical feasibility of UR-ddPCR by performing it intraoperatively for 13 glioma cases. We further combine UR-ddPCR measurements with UR-stimulated Raman histology intraoperatively to estimate tumor cell densities in addition to tumor cell percentages. We anticipate that UR-ddPCR, along with future refinements in assay instrumentation, will enable novel point-of-care diagnostics and the development of molecularly-guided surgeries that improve clinical outcomes.

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Section: Discussionmentioning

confidence: 99%

Ultra-Rapid Droplet Digital PCR Enables Intraoperative Tumor Quantification

Murphy,

Bianchini,

Smith

et al. 2024

Preprint

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“…Thus, the development of a portable integrated digital platform with easy-to-use workflows for automatic detection of target nucleic acids would greatly enhance digital CRISPR-based amplification-free analysis. Although some portable digital devices have been developed, they still require a microfluidic system and chips for sample partitioning, which can increase the cost and complexity of the device. − …”

Section: Discussionmentioning

confidence: 99%

DECODE: Contamination-Free Digital CRISPR Platform for Point-of-Care Detection of Viral DNA/RNA

Li,

Yin,

Zheng

et al. 2024

ACS Sens.

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Rapid and precise nucleic acid testing at the pointof-care (POC) is essential for effective screening and management of infectious diseases. Current polymerase-based molecular diagnostics often suffer from potential cross-contamination issues, particularly in POC settings. Here, we introduce DECODE, a contamination-free nucleic acid detection platform integrating digital microfluidics (DMF) for nucleic acid extraction and a digital CRISPR amplification-free assay for pathogen detection. The digital CRISPR assay demonstrates sensitivity, detecting target DNA and RNA in the reaction mixture at concentrations of 10 and 5 copies/μL, respectively. Leveraging DMF-extracted samples enhances the performance of the digital CRISPR amplification-free assay. DECODE offers a sample-to-result workflow of 75 min using compact devices. Validation studies using clinical samples confirm DECODE's robust performance, achieving 100% sensitivity and specificity in detecting HPV18 from cervical epithelial cells and influenza A from nasal swabs. DECODE represents a versatile, contamination-free detection platform poised to enhance integrated public health surveillance efforts.

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“…Additionally, the structure of the optical path and tubes enlarges the platform's dimensions, posing challenges for the design of the multi-sample parallel analysis platform. In the chip developed by Yulin Ren et al , 18 droplets are amplified within a flat chamber, followed by the capture of fluorescence intensity through direct imaging. This chip boasts a simple structure and high throughput.…”

Section: Introductionmentioning

confidence: 99%

A ddPCR platform based on a microfluidic chip with a dual-function flow-focusing structure for sample-to-result DNA quantification analysis

Zhang,

Wang,

Wang

et al. 2024

Lab Chip

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A three-in-one microfluidic droplet digital PCR platform for absolute quantitative analysis of DNA

Abstract: Droplet digital polymerase chain reaction (ddPCR) technology has found widespread applications in the ultrasensitive analysis of nucleic acids, where integrated ddPCR platforms with the capability of automated sample dispersion, followed...

Cited by 8 publications

References 27 publications

Ultra-Rapid Droplet Digital PCR Enables Intraoperative Tumor Quantification

Ultra-Rapid Droplet Digital PCR Enables Intraoperative Tumor Quantification

DECODE: Contamination-Free Digital CRISPR Platform for Point-of-Care Detection of Viral DNA/RNA

A ddPCR platform based on a microfluidic chip with a dual-function flow-focusing structure for sample-to-result DNA quantification analysis

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