Droplet digital PCR enabled by microfluidic impact printing for absolute gene quantification

Pan, Yang; Ma, Tuo; Meng, Qi; Mao, Yuxin; Chu, Kaiqin; Men, Yongfeng; Pan, Tingrui; Li, Baoqing; Chu, Jiaru

doi:10.1016/j.talanta.2019.120680

Cited by 33 publications

(16 citation statements)

References 48 publications

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“…applied a microfluidic impact printer technology to print well‐patterned nanoliter droplets on a hydrophobic surface. [ 23 ] After covering the droplet array with oil, the PCR amplification could be performed. The device revealed the different levels of p53 gene between colon cancer tissues and adjacent nontumorous tissues.…”

Section: Optofluidic Genetic Analysis For Biodiagnostic Applicationsmentioning

confidence: 99%

Emerging optofluidic technologies for biodiagnostic applications

Dai

et al. 2021

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The unprecedented global COVID‐19 pandemic strongly argues the critical need for innovative diagnostic tools meeting the requirement of test speed, accuracy, and throughput. Owing to the integration of optics and microfluidics technologies, optofluidic technology enables highly precise flow manipulation and highly sensitive signal detection at the microscale, thus offering the promising potential for developing biodiagnostic applications. Research toward this direction is fast growing into an emerging area. In this paper, we give an overview of emerging optofluidic technologies for biodiagnostic applications with the focus on three common types of biomarkers: nucleic acid, protein, and cell. We conclude by discussing the challenges, opportunities, and future perspectives of this field.

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Section: Optofluidic Genetic Analysis For Biodiagnostic Applicationsmentioning

confidence: 99%

Emerging optofluidic technologies for biodiagnostic applications

Dai

et al. 2021

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“…Digital PCR can achieve absolute quantification of target nucleic acids with high sensitivity, excellent precision, and superior resolution by dividing the sample into a large number of partitions, specifically amplifying the target nucleic acid sequence, and measuring the fluorescence signal at the end-point. 1–3 A variety of microfluidic methods with different physical designs and working principles have been developed to carry out digital PCR, including pneumatic pumping, 4,5 flow partitioning, 6–9 centrifugation, 10 self-digitization, 11,12 droplet printing, 13–15 microwell compartmentalization, 16 mechanical vibration 17 and the SlipChip. 18–20 Digital PCR is becoming an essential tool for quantitative nucleic acid analysis in both life science research and clinical molecular diagnostic fields.…”

Section: Introductionmentioning

confidence: 99%

Multiplex digital PCR with digital melting curve analysis on a self-partitioning SlipChip

Pan

et al. 2022

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“…The gold standard culture method and serological detection method are commonly used for the clinical detection of E. coli [8,10]. However, the operations of these methods are tedious and include pre-enrichment, selective plating, biochemical screening, and serological confirmation [11][12][13]. Generally, 2 ~ 3 days are needed to complete the detection of the target bacteria.…”

Section: Introductionmentioning

confidence: 99%

“…Enzyme-linked immunosorbent assay (ELISA) is a plate-based assay technique for detecting bacteria that has the advantages of fast detection speed [18], high throughput and good stability. However, the signal amplification of this method is minimal, and the choices for antibody labels are limited; therefore, the ELSA method cannot fully meet the needs of rapid detection of bacteria [13,19] Recently, the adenosine triphosphate (ATP) luminescent method was shown to be an alternative method for cell detection. In the presence of the catalyst enzyme, ATP reacts with luciferin to emit light.…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Microfluidics for the rapid detection of Escherichia coli O157:H7 using antibody-coated microspheres

Shopsin

Sun

et al. 2021

Bioengineered

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This study developed a novel method for the rapid detection of Escherichia coli (E. coli) O157:H7 on a microfluidic platform. First, the concentration of bacteria in a sample was determined with the adenosine triphosphate (ATP) method. Then, the specific detection of E. coli was achieved in a microfluidic chip by the immune-microsphere technique. The influences of the culture time, flow rate and capture time on the detection of the target bacteria were investigated systematically. Generally, with increasing capture time, more bacteria could be captured by the microspheres, which had a positive effect on bacterial detection. Furthermore, the sensitivity and specificity of the method were also tested. The results showed that this method could specifically detect E. coli with a sensitivity as high as 49.1 cfu/μL; the consumption of bacteria was 1 μL, and the reagent was at the microliter level. The testing time can be controlled within one and a half hours, and the cost of testing was approximately RMB 10. The method described in this article is simple and accurate and has great application value in bacterial detection for medical diagnostics.

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Droplet digital PCR enabled by microfluidic impact printing for absolute gene quantification

Cited by 33 publications

References 48 publications

Emerging optofluidic technologies for biodiagnostic applications

Emerging optofluidic technologies for biodiagnostic applications

Multiplex digital PCR with digital melting curve analysis on a self-partitioning SlipChip

Microfluidics for the rapid detection of Escherichia coli O157:H7 using antibody-coated microspheres

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