Development of a Low-Cost and Portable Real-Time PCR Machine for Developing Countries

Nguyen, Hong Ha; Tran, Viet; Ngo, Hoan T.

doi:10.1007/978-3-030-75506-5_6

Cited by 3 publications

(2 citation statements)

References 12 publications

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“…For isothermal temperature control, we employed a positive temperature coefficient (PTC) ceramic heater (Fig. S9) (Ngo et al, 2022), which has several advantages including fast warm-up, relatively uniform heating, and self-regulating, therefore being safe and energy efficient. Indeed, using the PTC ceramic heater, we achieved relatively uniform temperature across the heater surface (Fig.…”

Section: Compact Digital Assay Platformmentioning

confidence: 99%

Rapid and Portable Quantification of HIV RNA via a Smartphone-enabled Digital CRISPR Device and Deep Learning

Ngo

Akarapipad

Lee

et al. 2023

Preprint

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For the 28.2 million people in the world living with HIV/AIDS and receiving antiretroviral therapy, it is crucial to monitor their HIV viral loads with ease. To this end, rapid and portable diagnostic tools that can quantify HIV RNA are critically needed. We report herein a rapid and quantitative digital CRISPR-assisted HIV RNA detection assay that has been implemented within a portable smartphone-based device as a potential solution. Specifically, we first developed a fluorescence-based reverse transcription recombinase polymerase amplification (RT-RPA)-CRISPR assay for isothermally and rapidly detecting HIV RNA at 42 C in < 30 min. When realized within a commercial stamp-sized digital chip, this assay yields strongly fluorescent digital reaction wells corresponding to HIV RNA. The isothermal reaction condition and the strong fluorescence in the small digital chip unlock compact thermal and optical components in our device, allowing us to engineer a palm-size (70 by 115 by 80 mm) and lightweight (< 0.6 kg) device. Further leveraging the smartphone, we wrote a custom app to control the device, perform the digital assay, and acquire fluorescence images throughout the assay time. We additionally trained and verified a Deep Learning-based algorithm for analyzing fluorescence images and detecting strongly fluorescent digital reaction wells. Using our smartphone-enabled digital CRISPR device, we were able to detect 75 copies of HIV RNA in 15 min and demonstrate the potential of our device toward convenient monitoring of HIV viral loads and combating the HIV/AIDS epidemic.

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Section: Compact Digital Assay Platformmentioning

confidence: 99%

Rapid and Portable Quantification of HIV RNA via a Smartphone-enabled Digital CRISPR Device and Deep Learning

Ngo

Akarapipad

Lee

et al. 2023

Preprint

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show abstract

“…Among techniques for quantifying nucleic acid copy number in a single bulk reaction, real-time RT-PCR has better quantitative accuracy than isothermal techniques. , Nevertheless, real-time RT-PCR requires bulky and expensive instruments due to the need of thermal cycling and fluorescence readout . Several research groups have attempted to develop low-cost and portable real-time RT-PCR instruments for applications at the POC. − We previously reported a filtration-assisted HIV VL quantification method based upon a portable magnetofluidic real-time RT-PCR platform with a LOD of 1000 copies using 10 μL of whole blood, i.e., 100,000 copies/mL of blood and a good quantitative accuracy . However, the WHO defines ≥1000 HIV RNA copies/mL as the threshold for virological failure, while some other public health authorities use lower thresholds (e.g., ≥200 copies/mL). , Therefore, to be able to detect virological failure, a platform must achieve a LOD of 1000 HIV RNA copies/mL or lower.…”

Section: Introductionmentioning

confidence: 99%

Sensitive and Quantitative Point-of-Care HIV Viral Load Quantification from Blood Using a Power-Free Plasma Separation and Portable Magnetofluidic Polymerase Chain Reaction Instrument

et al. 2022

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Point-of-care (POC) HIV viral load (VL) tests are needed to enhance access to HIV VL testing in low-and middleincome countries (LMICs) and to enable HIV VL self-testing at home, which in turn have the potential to enhance the global management of the disease. While methods based on real-time reverse transcription-polymerase chain reaction (RT-PCR) are highly sensitive and quantitatively accurate, they often require bulky and expensive instruments, making applications at the POC challenging. On the other hand, although methods based on isothermal amplification techniques could be performed using lowcost instruments, they have shown limited quantitative accuracies, i.e., being only semiquantitative. Herein, we present a sensitive and quantitative POC HIV VL quantification method from blood that can be performed using a small power-free three-dimensionalprinted plasma separation device and a portable, low-cost magnetofluidic real-time RT-PCR instrument. The plasma separation device, which is composed of a plasma separation membrane and an absorbent material, demonstrated 96% plasma separation efficiency per 100 μL of whole blood. The plasma solution was then processed in a magnetofluidic cartridge for automated HIV RNA extraction and quantification using the portable instrument, which completed 50 cycles of PCR in 15 min. Using the method, we achieved a limit of detection of 500 HIV RNA copies/mL, which is below the World Health Organization's virological failure threshold, and a good quantitative accuracy. The method has the potential for sensitive and quantitative HIV VL testing at the POC and at home self-testing.

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A Precision, High Intensity and Programmable Current Power Supply for LED in Biomedical Applications

Ngoc Tran,

Thanh Nguyen,

Thi Thuy Le

et al. 2023

IFMBE Proceedings

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Development of a Low-Cost and Portable Real-Time PCR Machine for Developing Countries

Cited by 3 publications

References 12 publications

Rapid and Portable Quantification of HIV RNA via a Smartphone-enabled Digital CRISPR Device and Deep Learning

Rapid and Portable Quantification of HIV RNA via a Smartphone-enabled Digital CRISPR Device and Deep Learning

Sensitive and Quantitative Point-of-Care HIV Viral Load Quantification from Blood Using a Power-Free Plasma Separation and Portable Magnetofluidic Polymerase Chain Reaction Instrument

A Precision, High Intensity and Programmable Current Power Supply for LED in Biomedical Applications

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