An integrated lateral flow assay for effective DNA amplification and detection at the point of care

Choi, Jane Ru; Hu, Jie; Gong, Yan; Feng, Shangsheng; Abas, Wan Abu Bakar Wan; Murphy, Belinda; Xu, Feng

doi:10.1039/c5an02532j

Cited by 85 publications

(56 citation statements)

References 51 publications

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“…Meeting the emerging paradigm of medicine, in which pharmacogenomics and individualised theranostics are of increasing importance for patient stratification and avoidance of adverse drug effects, there is a clear need for rapid, inexpensive, highly sensitive and simple-to-use companion diagnostic tests for the qualitative/quantitative detection of nucleic acids9. Meeting this requirement, there are a large number of paper analytical devices (PAD) that have been developed for detection of PCR products using lateral flow assays.…”

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confidence: 99%

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Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay

Jauset‐Rubio

Svobodová

Mairal

et al. 2016

Sci Rep

139

113

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Sensitive, specific, rapid, inexpensive and easy-to-use nucleic acid tests for use at the point-of-need are critical for the emerging field of personalised medicine for which companion diagnostics are essential, as well as for application in low resource settings. Here we report on the development of a point-of-care nucleic acid lateral flow test for the direct detection of isothermally amplified DNA. The recombinase polymerase amplification method is modified slightly to use tailed primers, resulting in an amplicon with a duplex flanked by two single stranded DNA tails. This tailed amplicon facilitates detection via hybridisation to a surface immobilised oligonucleotide capture probe and a gold nanoparticle labelled reporter probe. A detection limit of 1 × 10−11 M (190 amol), equivalent to 8.67 × 105 copies of DNA was achieved, with the entire assay, both amplification and detection, being completed in less than 15 minutes at a constant temperature of 37 °C. The use of the tailed primers obviates the need for hapten labelling and consequent use of capture and reporter antibodies, whilst also avoiding the need for any post-amplification processing for the generation of single stranded DNA, thus presenting an assay that can facilely find application at the point of need.

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confidence: 99%

“…The use of LAMP combined with NALF has also been reported, but requires a post-LAMP denaturation step at 95 °C prior to detection9.…”

mentioning

confidence: 99%

Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay

Jauset‐Rubio

Svobodová

Mairal

et al. 2016

Sci Rep

139

113

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“…12 With advances in POC testing, [18][19][20][21][22] lateral flow assays (LFAs) hold tremendous potential to address the abovementioned challenges 23,24 due to the simple, affordable and user-friendly features. 25 However, the existing LFAs are associated with the limitation of poor sensitivity. 11,26 Several methods have been developed to enhance the sensitivity of LFAs, such as sample concentration, [27][28][29][30] thermal contrast, 31 fluidic control, [32][33][34] temperature-humidity technique, 35 probe-based signal enhancement, 23,36 enzyme-based signal amplification 37,38 and electrochemical devices-based enhancement.…”

Section: Introductionmentioning

confidence: 99%

Improved LFIAs for highly sensitive detection of BNP at point-of-care

Gong¹,

Hu²,

Choi³

et al. 2017

IJN

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Heart failure (HF) has become a major cause of morbidity and mortality with a significant global economic burden. Although well-established clinical tests could provide early diagnosis, access to these tests is limited in developing countries, where a relatively higher incidence of HF is present. This has prompted an urgent need for developing a cost-effective, rapid and robust diagnostic tool for point-of-care (POC) detection of HF. Lateral flow immunoassay (LFIA) has found widespread applications in POC diagnostics. However, the low sensitivity of LFIA limits its ability to detect important HF biomarkers (e.g., brain natriuretic peptide [BNP]) that are normally present in low concentration in blood. To address this issue, we developed an improved LFIA by optimizing the gold nanoparticle (GNP)–antibody conjugate conditions (e.g., the conjugate pH and the amount of added antibody), the diameter of GNP and the concentration of antibody embedded on the test line and modifying the structure of test strip. Through these improvements, the proposed test strip enabled the detection of BNP down to 0.1 ng/mL within 10–15 min, presenting ~15-fold sensitivity enhancement over conventional lateral flow assay. We also successfully applied our LFIA in the analysis of BNP in human serum samples, highlighting its potential use for clinical assessment of HF. The developed LFIA for BNP could rapidly rule out HF with the naked eye, offering tremendous potential for POC test and personalized medicine.

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“…5E) was utilized to detect DENV (Choi et al, 2016). The nanoparticles with ssDNA detector probe are typically prepared and assembled on the LFS membrane.…”

Section: Detection Methods For Locmentioning

confidence: 99%

“…A fully integrated paper-based LAMP device with an LFS layer (Fig. (E) LFS-based NA amplification detection (Choi et al, 2016). They immobilized gold nanoparticles and detect probe conjugates on a nitrocellulose membrane for the LFS layer to capture target amplicons.…”

Section: Detection Methods For Locmentioning

confidence: 99%

Recent advances in lab-on-a-chip technologies for viral diagnosis

Zhu

Fohlerová

Pekárek

et al. 2020

Biosensors and Bioelectronics

190

146

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A B S T R A C TThe global risk of viral disease outbreaks emphasizes the need for rapid, accurate, and sensitive detection techniques to speed up diagnostics allowing early intervention. An emerging field of microfluidics also known as the lab-on-a-chip (LOC) or micro total analysis system includes a wide range of diagnostic devices. This review briefly covers both conventional and microfluidics-based techniques for rapid viral detection. We first describe conventional detection methods such as cell culturing, immunofluorescence or enzyme-linked immunosorbent assay (ELISA), or reverse transcription polymerase chain reaction (RT-PCR). These methods often have limited speed, sensitivity, or specificity and are performed with typically bulky equipment. Here, we discuss some of the LOC technologies that can overcome these demerits, highlighting the latest advances in LOC devices for viral disease diagnosis. We also discuss the fabrication of LOC systems to produce devices for performing either individual steps or virus detection in samples with the sample to answer method. The complete system consists of sample preparation, and ELISA and RT-PCR for viral-antibody and nucleic acid detection, respectively. Finally, we formulate our opinions on these areas for the future development of LOC systems for viral diagnostics.

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An integrated lateral flow assay for effective DNA amplification and detection at the point of care

Cited by 85 publications

References 51 publications

Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay

Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay

Improved LFIAs for highly sensitive detection of BNP at point-of-care

Recent advances in lab-on-a-chip technologies for viral diagnosis

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