Ultra-rapid real-time microfluidic RT-PCR instrument for nucleic acid analysis

Nouwairi, Renna; Cunha, Larissa L.; Turiello, Rachelle; Scott, Orion N.; Hickey, Jeff; Thompson, Scott; Knowles, Stuart; Chapman, Jeff; Landers, James P.

doi:10.1039/d2lc00495j

Cited by 20 publications

(13 citation statements)

References 29 publications

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“…In fact, it is often observed in fast PCR that amplification efficiency or sensitivity is sacrificed in favor of fast results [ 9 ]. In many applications, a binary (yes/no) answer is sought whether the target is present or not, especially in clinical diagnostics [ 38 ].…”

Section: Resultsmentioning

confidence: 99%

Ultrafast Microfluidic PCR Thermocycler for Nucleic Acid Amplification

Huang

et al. 2023

Micromachines

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The polymerase chain reaction (PCR) is essential in nucleic acid amplification tests and is widely used in many applications such as infectious disease detection, tumor screening, and food safety testing; however, most PCR devices have inefficient heating and cooling ramp rates for the solution, which significantly limit their application in special scenarios such as hospital emergencies, airports, and customs. Here, we propose a temperature control strategy to significantly increase the ramp rates for the solution temperature by switching microfluidic chips between multiple temperature zones and excessively increasing the temperature difference between temperature zones and the solution; accordingly, we have designed an ultrafast thermocycler. The results showed that the ramp rates of the solution temperature are a linear function of temperature differences within a range, and a larger temperature difference would result in faster ramp rates. The maximum heating and cooling ramp rates of the 25 μL solution reached 24.12 °C/s and 25.28 °C/s, respectively, and the average ramp rate was 13.33 °C/s, 6–8 times higher than that of conventional commercial PCR devices. The thermocycler achieved 9 min (1 min pre-denaturation + 45 PCR cycles) ultrafast nucleic acid amplification, shortening the time by 92% compared to the conventional 120 min nucleic acid amplification, and has the potential to be used for rapid nucleic acid detection.

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Section: Resultsmentioning

confidence: 99%

Ultrafast Microfluidic PCR Thermocycler for Nucleic Acid Amplification

Huang

et al. 2023

Micromachines

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“…Microfluidics combined with nucleic acid amplification can lead to a more sensitive test result and make it a powerful tool for clinical microbiology testing. In recent years, various microfluidic devices based on nucleic acid amplification have emerged for the rapid and accurate detection of DNA and RNA [ [56] , [57] , [58] ]. In general, nucleic acid amplification-based techniques are classified as PCR and isothermal nucleic acid amplification.…”

Section: Microfluidic Chip and On-chip Nucleic Acid Detectionmentioning

confidence: 99%

Recent advancements in nucleic acid detection with microfluidic chip for molecular diagnostics

Wang

et al. 2023

TrAC Trends in Analytical Chemistry

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“…From the heat transfer perspective, some parameters in the microfluidic chip design may affect the PCR efficiency, such as the sample volume within the chip's PCR chamber, the film's thickness on both sides and the specific surface area for heat transfer. 23,40,41 This will be detailed in the Numerical simulation of thermodynamics section. In addition, the PC film used in the PCR chamber is only about 0.1 mm thick and difficult to shape.…”

Section: Overview and Composition Of The Systemmentioning

confidence: 99%

“…22 Renna L. Nouwairi et al designed a thermal cycler that ensures swift and effective thermal energy transmission through close contact between the clamped Peltier and the PCR chamber, completing 40 PCR cycles in 10 min, also equipped with a photoelectric detection interface, which delivers sensitivity and efficiency on par with commercial instruments. 23 Jiyong Cheong et al designed a portable device for nucleic acid detection based on plasma heating of magnetic plasma nanoparticles and in situ fluorescence detection after magnetic nanoparticle removal, with a rapid heating rate of 13.17 °C s −1 and a cooling rate of 4.94 °C s −1 . The RNA of SARS-CoV-2 was detected within 17 min.…”

Section: Introductionmentioning

confidence: 99%