Rapid developments in lateral flow immunoassay for nucleic acid detection

Zheng, Chengshi; Wang, Kan; Zheng, Wei; Cheng, Yuemeng; Li, Tangan; Cao, Bo; Jin, Qinghui; Cui, Daxiang

doi:10.1039/d0an02150d

Cited by 65 publications

(49 citation statements)

References 118 publications

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“…The enzyme’s strong strand displacement activity overcomes the necessity of temperature cycling of RT-PCR. Exploiting the reverse transcriptase property of the enzyme, RT-LAMP is also a suitable amplification method for RNA [ 16 , 17 ]. Moreover, the RT-LAMP product may also be read out simply by using an immunochromatographic, paper-based lateral flow assays (LFA) when certain modifications are considered [ 18 , 19 ].…”

Section: Introductionmentioning

confidence: 99%

Lateral flow–based nucleic acid detection of SARS-CoV-2 using enzymatic incorporation of biotin-labeled dUTP for POCT use

et al. 2022

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The degree of detrimental effects inflicted on mankind by the COVID-19 pandemic increased the need to develop ASSURED (Affordable, Sensitive, Specific, User-friendly, Rapid and Robust, Equipment-free, and Deliverable) POCT (point of care testing) to overcome the current and any future pandemics. Much effort in research and development is currently advancing the progress to overcome the diagnostic pressure built up by emerging new pathogens. LAMP (loop-mediated isothermal amplification) is a well-researched isothermal technique for specific nucleic acid amplification which can be combined with a highly sensitive immunochromatographic readout via lateral flow assays (LFA). Here we discuss LAMP-LFA robustness, sensitivity, and specificity for SARS-CoV-2 N-gene detection in cDNA and clinical swab-extracted RNA samples. The LFA readout is designed to produce highly specific results by incorporation of biotin and FITC labels to 11-dUTP and LF (loop forming forward) primer, respectively. The LAMP-LFA assay was established using cDNA for N-gene with an accuracy of 95.65%. To validate the study, 82 SARS-CoV-2-positive RNA samples were tested. Reverse transcriptase (RT)-LAMP-LFA was positive for the RNA samples with an accuracy of 81.66%; SARS-CoV-2 viral RNA was detected by RT-LAMP-LFA for as low as CT-33. Our method reduced the detection time to 15 min and indicates therefore that RT-LAMP in combination with LFA represents a promising nucleic acid biosensing POCT platform that combines with smartphone based semi-quantitative data analysis. Graphical abstract

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Section: Introductionmentioning

confidence: 99%

Lateral flow–based nucleic acid detection of SARS-CoV-2 using enzymatic incorporation of biotin-labeled dUTP for POCT use

et al. 2022

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“…The amplicons can be detected in different ways: visualization by gel electrophoresis, by real-time detection using fluorescent dye or probes, and by specific and affine recognition of tag-labeled amplicons in lateral flow test (LFT) [ 1 , 3 ]. The LFT is the most popular tool for sensitive and rapid detection of amplicons that is effective in resource-limited conditions [ 4 ]. To realize RPA-LFT, two different specific tags should be included in the amplicons during the RPA process.…”

Section: Introductionmentioning

confidence: 99%

Recombinase Polymerase Amplification Assay with and without Nuclease-Dependent-Labeled Oligonucleotide Probe

Иванов

Safenkova

Zherdev

et al. 2021

IJMS

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The combination of recombinase polymerase amplification (RPA) and lateral flow test (LFT) is a strong diagnostic tool for rapid pathogen detection in resource-limited conditions. Here, we compared two methods generating labeled RPA amplicons following their detection by LFT: (1) the basic one with primers modified with different tags at the terminals and (2) the nuclease-dependent one with the primers and labeled oligonucleotide probe for nuclease digestion that was recommended for the high specificity of the assay. Using both methods, we developed an RPA-LFT assay for the detection of worldwide distributed phytopathogen—alfalfa mosaic virus (AMV). A forward primer modified with fluorescein and a reverse primer with biotin and fluorescein-labeled oligonucleotide probe were designed and verified by RPA. Both labeling approaches and their related assays were characterized using the in vitro-transcribed mRNA of AMV and reverse transcription reaction. The results demonstrated that the RPA-LFT assay based on primers-labeling detected 103 copies of RNA in reaction during 30 min and had a half-maximal binding concentration 22 times lower than probe-dependent RPA-LFT. The developed RPA-LFT was successfully applied for the detection of AMV-infected plants. The results can be the main reason for choosing simple labeling with primers for RPA-LFT for the detection of other pathogens.

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“…Although these methods detect target nucleic acids via hybridization to complementary sequences, their binding principles are different as NALF directly detects DNA exploiting capture and labeled reporter oligonucleotide probes, whereas NALFIA detects hapten-labeled DNA using capture and labeled reporter antibodies or streptavidin ( Jauset-Rubio et al, 2016b ). A typical workflow of lateral flow nucleic acid tests comprises nucleic acid extraction from biological samples, nucleic acid amplification, and the detection of an analyte through LFAs using labeled color-changing agents ( Zheng C. et al, 2021 ).…”

Section: Paper-based Sars-cov-2 Diagnostic Methodsmentioning

confidence: 99%

Paper-Based Point-of-Care Testing of SARS-CoV-2

Yuan

Sun

Tian

et al. 2021

Front. Bioeng. Biotechnol.

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The COVID-19 pandemic has resulted in significant global social and economic disruption. The highly transmissive nature of the disease makes rapid and reliable detection critically important. Point-of-care (POC) tests involve performing diagnostic tests outside of a laboratory that produce a rapid and reliable result. It therefore allows the diagnostics of diseases at or near the patient site. Paper-based POC tests have been gaining interest in recent years as they allow rapid, low-cost detection without the need for external instruments. In this review, we focus on the development of paper-based POC devices for the detection of SARS-CoV-2. The review first introduces the principles of detection methods that are available to paper-based devices. It then summarizes the state-of-the-art paper devices and their analytical performances. The advantages and drawbacks among methods are also discussed. Finally, limitations of the existing devices are discussed, and prospects are given with the hope to identify research opportunities and directions in the field. We hope this review will be helpful for researchers to develop a clinically useful and economically efficient paper-based platform that can be used for rapid, accurate on-site diagnosis to aid in identifying acute infections and eventually contain the COVID-19 pandemic.

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Rapid developments in lateral flow immunoassay for nucleic acid detection

Abstract: Recently, lateral flow assay (LFA) for nucleic acid detection has drawn increasing attention in the point-of-care testing fields. Thanks to its rapidity, easy implementation, and low equipment requirement, it is...

Cited by 65 publications

References 118 publications

Lateral flow–based nucleic acid detection of SARS-CoV-2 using enzymatic incorporation of biotin-labeled dUTP for POCT use

Lateral flow–based nucleic acid detection of SARS-CoV-2 using enzymatic incorporation of biotin-labeled dUTP for POCT use

Recombinase Polymerase Amplification Assay with and without Nuclease-Dependent-Labeled Oligonucleotide Probe

Paper-Based Point-of-Care Testing of SARS-CoV-2

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