Detection of influenza virus using a lateral flow immunoassay for amplified DNA by a microfluidic RT-PCR chip

Nagatani, Naoki; Yamanaka, Keiichiro; Ushijima, Hiromi; Koketsu, Ritsuko; Sasaki, Tadahiro; Ikuta, Kazuyoshi; Saito, Masato; Miyahara, Tetsuhiro; Tamiya, Eiichi

doi:10.1039/c2an16294f

Cited by 38 publications

(37 citation statements)

References 34 publications

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“…Whilst PCR is the most commonly reported method of amplification combined with lateral flow192021, there are an increasing number of reports combining isothermal amplification with lateral flow detection222324252627282930, moving nearer to achieving ASSURED devices that can truly be used at the point-of-need.…”

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

144

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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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

144

113

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“…22,23,27,[38][39][40][41][42] As the fluid moves through the serpentine channel, the fluid experiences different temperatures in different areas of the chip (Fig. 22,23,27,[38][39][40][41][42] As the fluid moves through the serpentine channel, the fluid experiences different temperatures in different areas of the chip (Fig.…”

Section: Continuous Flowmentioning

confidence: 99%

“…26). 26,42,84 The conjugate pad stores a conjugate, described below in more detail. The sample then migrates, by capillary action, through the conjugate pad, down the nitrocellulose membrane, and into the absorbent pad.…”

Section: Lateral Flowmentioning

confidence: 99%

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Portable nucleic acid thermocyclers

2013

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A nucleic acid thermal cycler is considered to be portable if it is under ten pounds, easily carried by one individual, and battery powered. Nucleic acid amplification includes both polymerase chain reaction (e.g. PCR, RT-PCR) and isothermal amplification (e.g. RPA, HDA, LAMP, NASBA, RCA, ICAN, SMART, SDA). There are valuable applications for portable nucleic acid thermocyclers in fields that include clinical diagnostics, biothreat detection, and veterinary testing. A system that is portable allows for the distributed detection of targets at the point of care and a reduction of the time from sample to answer. The designer of a portable nucleic acid thermocycler must carefully consider both thermal control and the detection of amplification. In addition to thermal control and detection, the designer may consider the integration of a sample preparation subsystem with the nucleic acid thermocycler. There are a variety of technologies that can achieve accurate thermal control and the detection of nucleic acid amplification. Important evaluation criteria for each technology include maturity, power requirements, cost, sensitivity, speed, and manufacturability. Ultimately the needs of a particular market will lead to user requirements that drive the decision between available technologies.

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“…On the basis of PCR, scientists have developed a series of technologies, such as quantitative real-time PCR (Demidenko and Penin, 2012), multiplex PCR (Becker et al, 2012), chip PCR , microfluidic PCR (Nagatani et al, 2012), and isothermal PCR (Paris et al, 2011). Loop-mediated isothermal amplification (LAMP) was invented by Notomi et al (2000), which is characterized as rapid and of high specificity and sensitivity.…”

Section: Introductionmentioning

confidence: 99%

DNA detection of Clostridium difficile infection based on real-time resistance measurement

Liu¹,

Jiang²,

Xiang³

et al. 2013

Genet. Mol. Res.

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ABSTRACT. We used a newly developed electrochemical method, real-time resistance measurement, based on loop-mediated isothermal amplification (LAMP), with real-time resistance monitoring and derivative analysis. DNA extracted from specimens was amplified through LAMP reaction. The 2 products of LAMP, DNA and pyrophosphate, both are negative ions; they combine with positive dye (crystal violet) and positive ions (Mg 2+ ), which leads to an increase in the resistivity of the reaction liquid. The changes of resistivity were measured in real-time with a specially designed resistance electrode, to detect Clostridium difficile DNA. We found that electrochemical detection of C. difficile could be completed in 0.5-1 h, with a detection limit of 10 2 CFU/mL, with high accuracy (95.0%), sensitivity (91.1%), and specificity (97.3%) compared to PCR methods. C. difficile is commonly associated with antibioticinduced diarrhea. Due to the difficulty in performing anaerobic culture and cytotoxicity neutralization assays, a simple, rapid, sensitive, and accurate method is preferred. We conclude that realtime resistance measurement is a rapid, sensitive, and stable method for the diagnosis of C. difficile infection that could be applied to gene chips and pocket instruments.

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Detection of influenza virus using a lateral flow immunoassay for amplified DNA by a microfluidic RT-PCR chip

Abstract: Influenza virus RNA was amplified by a continuous-flow polydimethylsiloxane microfluidic RT-PCR chip within 15-20 min. The amplified influenza virus RNA was observed with the naked eye, as the red color at the test line, using a lateral flow immunoassay within 1 min.

Cited by 38 publications

References 34 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

Portable nucleic acid thermocyclers

DNA detection of Clostridium difficile infection based on real-time resistance measurement

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