Microfluidic Nano-Scale qPCR Enables Ultra-Sensitive and Quantitative Detection of SARS-CoV-2

Xie, Xin; Gjorgjieva, Tamara; Attieh, Zaynoun; Dieng, Mame Massar; Arnoux, Marc; Khair, Mostafa; Moussa, Yasmine; Jallaf, Fatima Al; Rahiman, Nabil; Jackson, Christopher; Messery, Lobna El; Pamplona, Khristine; Victoria, Zyrone; Zafar, Mohammed; Ali, Raghib; Piano, Fabio; Gunsalus, Kristin C.; Idaghdour, Youssef

doi:10.3390/pr8111425

Cited by 20 publications

(23 citation statements)

References 28 publications

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“…Xie et al . utilized a nano-scale RT-qPCR system on the microfluidic chip, which is one of the three-step approaches to detect SARS-CoV-2 and achieved the result of reduced False-Negative rate with low viral loads with higher sensitivity than the standard RT-PCR method [ 194 ]. Tarim et al .…”

Section: Micro/nanotechnology-based Diagnosis Methodsmentioning

confidence: 99%

Micro/nanotechnology-inspired rapid diagnosis of respiratory infectious diseases

et al. 2021

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Humans have suffered from a variety of infectious diseases since a long time ago, and now a new infectious disease called COVID-19 is prevalent worldwide. The ongoing COVID-19 pandemic has led to research of the effective methods of diagnosing respiratory infectious diseases, which are important to reduce infection rate and help the spread of diseases be controlled. The onset of COVID-19 has led to the further development of existing diagnostic methods such as polymerase chain reaction, reverse transcription polymerase chain reaction, and loop-mediated isothermal amplification. Furthermore, this has contributed to the further development of micro/nanotechnology-based diagnostic methods, which have advantages of high-throughput testing, effectiveness in terms of cost and space, and portability compared to conventional diagnosis methods. Micro/nanotechnology-based diagnostic methods can be largely classified into (1) nanomaterials-based, (2) micromaterials-based, and (3) micro/nanodevice-based. This review paper describes how micro/nanotechnologies have been exploited to diagnose respiratory infectious diseases in each section. The research and development of micro/nanotechnology-based diagnostics should be further explored and advanced as new infectious diseases continue to emerge. Only a handful of micro/nanotechnology-based diagnostic methods has been commercialized so far and there still are opportunities to explore.

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Section: Micro/nanotechnology-based Diagnosis Methodsmentioning

confidence: 99%

Micro/nanotechnology-inspired rapid diagnosis of respiratory infectious diseases

et al. 2021

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“…Fluorescence is the most extensively used approach due to its high sensitivity and abundance of fluorophores and labeling chemistries [ 80 ]. A quantitative nanofluidic assay based on qPCR developed by Fassy et al, three-step microfluidic nano-scale qPCR based on a microfluidic chip designed by Xie et al, microfluidic chip PCR technology developed by Francesca Dragoni et al, microfluidic disc-direct RT-qPCR assay developed by Ji M et al, portable MiDAS for SARS-CoV-2 nucleic acids detection developed by Yang, J et al, detect SARS-CoV-2 nucleic acids by the fluorescence-based detector ( Table 3 ) [ 81 , 82 , 83 , 84 , 85 ].…”

Section: Microfluidics In Sars-cov-2mentioning

confidence: 99%

“…Henceforth, timely diagnosis is required to avoid such outcomes [ 89 ]. To address the issue of false-negative results while simultaneously being able to detect low viral loads that are missed in the RT-PCR technique, Xie et al developed a three-step microfluidic qPCR method that incorporates reverse transcription, targeted cDNA pre-amplification, which enables detection below 1 copy/µL and nanoscale PCR methods [ 82 ]. Fassy et al also included the pre-amplification step enabling the detection of seven transcript copies per reaction for N genes.…”

Section: Microfluidics In Sars-cov-2mentioning

confidence: 99%

Microfluidics Technology in SARS-CoV-2 Diagnosis and Beyond: A Systematic Review

Jamiruddin

Meghla

Islam

et al. 2022

Life

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With the progression of the COVID-19 pandemic, new technologies are being implemented for more rapid, scalable, and sensitive diagnostics. The implementation of microfluidic techniques and their amalgamation with different detection techniques has led to innovative diagnostics kits to detect SARS-CoV-2 antibodies, antigens, and nucleic acids. In this review, we explore the different microfluidic-based diagnostics kits and how their amalgamation with the various detection techniques has spearheaded their availability throughout the world. Three other online databases, PubMed, ScienceDirect, and Google Scholar, were referred for articles. One thousand one hundred sixty-four articles were determined with the search algorithm of microfluidics followed by diagnostics and SARS-CoV-2. We found that most of the materials used to produce microfluidics devices were the polymer materials such as PDMS, PMMA, and others. Centrifugal force is the most commonly used fluid manipulation technique, followed by electrochemical pumping, capillary action, and isotachophoresis. The implementation of the detection technique varied. In the case of antibody detection, spectrometer-based detection was most common, followed by fluorescence-based as well as colorimetry-based. In contrast, antigen detection implemented electrochemical-based detection followed by fluorescence-based detection, and spectrometer-based detection were most common. Finally, nucleic acid detection exclusively implements fluorescence-based detection with a few colorimetry-based detections. It has been further observed that the sensitivity and specificity of most devices varied with implementing the detection-based technique alongside the fluid manipulation technique. Most microfluidics devices are simple and incorporate the detection-based system within the device. This simplifies the deployment of such devices in a wide range of environments. They can play a significant role in increasing the rate of infection detection and facilitating better health services.

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“…Polymerase chain reaction (PCR) is a well-established technique that is widely used in medical, molecular biology and synthetic biology settings for amplifying DNA segments. Most recently, reverse transcriptase PCR (RT-PCR) has seen some increased popularity due to its use as a diagnostic mechanism in the detection of SARS-CoV-2 [ 6 ]. PCR is a technique that can be used to amplify the desired segment of DNA or RNA, such as SARS-CoV-2 viral RNA [ 7 ].…”

Section: Introductionmentioning

confidence: 99%

Integrated Microfluidic-Based Platforms for On-Site Detection and Quantification of Infectious Pathogens: Towards On-Site Medical Translation of SARS-CoV-2 Diagnostic Platforms

Escobar

Chiu

et al. 2021

Micromachines

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The rapid detection and quantification of infectious pathogens is an essential component to the control of potentially lethal outbreaks among human populations worldwide. Several of these highly infectious pathogens, such as Middle East respiratory syndrome (MERS) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), have been cemented in human history as causing epidemics or pandemics due to their lethality and contagiousness. SARS-CoV-2 is an example of these highly infectious pathogens that have recently become one of the leading causes of globally reported deaths, creating one of the worst economic downturns and health crises in the last century. As a result, the necessity for highly accurate and increasingly rapid on-site diagnostic platforms for highly infectious pathogens, such as SARS-CoV-2, has grown dramatically over the last two years. Current conventional non-microfluidic diagnostic techniques have limitations in their effectiveness as on-site devices due to their large turnaround times, operational costs and the need for laboratory equipment. In this review, we first present criteria, both novel and previously determined, as a foundation for the development of effective and viable on-site microfluidic diagnostic platforms for several notable pathogens, including SARS-CoV-2. This list of criteria includes standards that were set out by the WHO, as well as our own “seven pillars” for effective microfluidic integration. We then evaluate the use of microfluidic integration to improve upon currently, and previously, existing platforms for the detection of infectious pathogens. Finally, we discuss a stage-wise means to translate our findings into a fundamental framework towards the development of more effective on-site SARS-CoV-2 microfluidic-integrated platforms that may facilitate future pandemic diagnostic and research endeavors. Through microfluidic integration, many limitations in currently existing infectious pathogen diagnostic platforms can be eliminated or improved upon.

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Microfluidic Nano-Scale qPCR Enables Ultra-Sensitive and Quantitative Detection of SARS-CoV-2

Cited by 20 publications

References 28 publications

Micro/nanotechnology-inspired rapid diagnosis of respiratory infectious diseases

Micro/nanotechnology-inspired rapid diagnosis of respiratory infectious diseases

Microfluidics Technology in SARS-CoV-2 Diagnosis and Beyond: A Systematic Review

Integrated Microfluidic-Based Platforms for On-Site Detection and Quantification of Infectious Pathogens: Towards On-Site Medical Translation of SARS-CoV-2 Diagnostic Platforms

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