Isothermal Target and Signaling Probe Amplification Method, Based on a Combination of an Isothermal Chain Amplification Technique and a Fluorescence Resonance Energy Transfer Cycling Probe Technology

Jung, Cheulhee; Chung, Jin; Kim, Un Ok; Kim, Min Hwan; Park, Hyun Gyu

doi:10.1021/ac100606m

Cited by 47 publications

(25 citation statements)

References 27 publications

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“…Recently, Jung et al (Jung et al 2010) developed a new highly sensitive and specific isothermal amplification and detection system called isothermal target and probe amplification (iTPA) by employing DNA-RNA-DNA chimeric primers and a FRET probe. The iTPA reaction is done under isothermal conditions between 55 and 65°C using a simple incubator such as a water bath or block heater which is sufficient for amplification.…”

Section: Rapid and Simple Detection Of Inva Gene In Salmonella Spp Bmentioning

confidence: 99%

Studies on PCR-Based Rapid Detection Systems for Salmonella spp.

Kim¹

2012

Salmonella - A Dangerous Foodborne Pathogen

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Section: Rapid and Simple Detection Of Inva Gene In Salmonella Spp Bmentioning

confidence: 99%

Studies on PCR-Based Rapid Detection Systems for Salmonella spp.

Kim¹

2012

Salmonella - A Dangerous Foodborne Pathogen

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“…A DNA-RNA-DNA chimeric primer set and a further outer primer set for strand displacement are used. RNase H cuts the RNA, anneals to the single strand and a strand-displacing polymerase generates the amplicon [191].…”

Section: Various Other Methodsmentioning

confidence: 99%

Isothermal Amplification and Quantification of Nucleic Acids and its Use in Microsystems

Tröger¹,

Niemann²,

Gärtig³

et al. 2015

J Nanomed Nanotechnol

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Nucleic acid amplification technologies (NAATs) offer the most sensitive tests in the clinical laboratory. These techniques are used as a powerful tool for screening and diagnosis of infectious diseases. Isothermal methods, as an alternative to polymerase chain reaction (PCR), require no thermocycling machine and can mostly be performed with reduced time, high throughput, and accurate and reliable results.However, current molecular diagnostic approaches generally need manual analysis by qualified and experienced personal which is a highly complex, time-consuming and labor-intensive task. Thus, the demand for simpler, miniaturized systems and assays for pathogen detection is steadily increasing. Microfluidic platforms and lab-on-a-chip devices have many advantages such as small sample volume, portability and rapid detection time and enable point-of-care diagnosis.In this article, we review several isothermal amplification methods and their implementation in microsystems in relation to quantification of nucleic acids. miniaturized systems can be diverse. The majority of publications are focussed on clinical diagnostics including foodborne organisms for environmental monitoring [10][11][12]. In case of an infectious disease, cultivation and phenotypic characterization are still the standard methods of microbiologists which need days up to weeks to obtain a diagnostic result. Hence, scientists started to make use of nucleic acid amplification methods to accelerate the analysis. The most widespread and well known technique for this purpose is the polymerase chain reaction (PCR) [10,13], which exploits the activity of the DNA polymerase. The method is based on a thermal cycling process consisting of repeated cycles of heating and cooling steps allowing for DNA melting, sequence specific primer binding and enzymatic amplification of the target nucleic acid. The quantitative assessment of target copies is possible by using a real-time PCR approach, where a concentration dependent signal (e.g. fluorescence) increases over time [14]. The latter enables the user to easily assess the pathogen load of patient samples [15][16][17][18]. Since the first example of a PCR on a miniaturized system in 1994 [19,20], numerous devices were presented, which integrate not just the amplification, but also sample preparation and detection steps [9,[21][22][23][24][25][26][27][28]. Just a few of those microsystems have reached the market so far, but some have shown a fascinating potential to replace the classical laboratory-oriented state-of-the art [29][30][31][32][33]. setups has been shown to obtain a similar result [34]. A smaller heat capacity allows for rapid changes in temperature beneficial for the PCR time as well as a higher parallelism of multiple genetic samples [34].However, a major drawback of the integration of PCR to microsystems is the necessity of sophisticated instrumentation. The reaction requires a thermal cycling instrumentation, space and considerable expertise [35]. Therefore, isothermal reactions for the ta...

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confidence: 99%

“…Isothermal amplification strategies such as recombinase polymerase amplification have been introduced as alternatives (2)(3)(4). Diagnostic applications include microRNA profiling, gene expression analysis, pathogen quantification, and cancer diagnostics (2,3 ). There is an ongoing development of novel devices on the basis of capillary systems, microfluidics, point-of-care testing, microbeads, and related technologies (6,(7)(8)(9)(10)(11)(12).…”

confidence: 99%

Impact of Smoothing on Parameter Estimation in Quantitative DNA Amplification Experiments

et al. 2015

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BACKGROUND: Quantification cycle (Cq) and amplification efficiency (AE) are parameters mathematically extracted from raw data to characterize quantitative PCR (qPCR) reactions and quantify the copy number in a sample. Little attention has been paid to the effects of preprocessing and the use of smoothing or filtering approaches to compensate for noisy data. Existing algorithms largely are taken for granted, and it is unclear which of the various methods is most informative. We investigated the effect of smoothing and filtering algorithms on amplification curve data.

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Isothermal Target and Signaling Probe Amplification Method, Based on a Combination of an Isothermal Chain Amplification Technique and a Fluorescence Resonance Energy Transfer Cycling Probe Technology

Cited by 47 publications

References 27 publications

Studies on PCR-Based Rapid Detection Systems for Salmonella spp.

Studies on PCR-Based Rapid Detection Systems for Salmonella spp.

Isothermal Amplification and Quantification of Nucleic Acids and its Use in Microsystems

Impact of Smoothing on Parameter Estimation in Quantitative DNA Amplification Experiments

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