An integrated microfluidic loop-mediated-isothermal-amplification system for rapid sample pre-treatment and detection of viruses

Wang, Chih‐Hung; Lien, Kang Yi; Wang, Ting Yu; Chen, Tzong Yueh; Lee, Gwo‐Bin

doi:10.1016/j.bios.2010.08.083

Cited by 96 publications

(59 citation statements)

References 29 publications

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Section: Strategies To Miniaturize Isothermal Amplification Reactionsmentioning

confidence: 99%

“…The HBV template could be detected by an SPR-LAMP system in 17 minutes even at the detection limit of 2 fg/mL. Several LAMP systems of on-chip detection modules were also combined with heating modules [52][53][54]. For the purpose of complete automatic control of the assay, Wang et al integrated sample pretreatment into the microfluidic LAMP device by using magnetic beads and a specially designed isolation membrane [53,54].…”

Section: Strategies To Miniaturize Isothermal Amplification Reactionsmentioning

confidence: 99%

“…Thus, many microsystems that combined LAMP with different detection methods, newly designed lab-on-chips, microfluidic cartridges and automated techniques were reported. In many studies, the LAMP amplification signals were already detected after 15-40 minutes after the thermal reaction started [50][51][52][53][54].The preferred and the most sensitive method for detection is based on fluorescence (with SYBR green, SYTO green or EvaGreen as fluorescent dyes) that can be detected as early as 15 minutes after the thermal reaction starts. For calcein, a limit of detection (LOD) of about 270 copies/µL was described [55].…”

mentioning

confidence: 99%

“…Several LAMP systems of on-chip detection modules were also combined with heating modules [52][53][54]. For the purpose of complete automatic control of the assay, Wang et al integrated sample pretreatment into the microfluidic LAMP device by using magnetic beads and a specially designed isolation membrane [53,54]. The entire diagnostic protocol including sample pre-treatment, LAMP reaction and optical detection was automatically completed in 60 minutes, which was twice as fast as with conventional devices [53].…”

mentioning

confidence: 99%

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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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Section: Strategies To Miniaturize Isothermal Amplification Reactionsmentioning

confidence: 99%

Section: Strategies To Miniaturize Isothermal Amplification Reactionsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

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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show abstract

“…Amplification of RNA viruses, as well as the more traditional single chamber for aqueous RT-PCR, has been achieved using both continuous flow 15 and solid-phase microarrays. 16 Isothermal techniques, such as nucleic acid sequence-based amplification 17 and loop-mediated isothermal amplification, 18 have also been successfully used.…”

mentioning

confidence: 99%

Integrated RNA extraction and RT-PCR for semi-quantitative gene expression studies on a microfluidic device

Shaw

Hughes

Dyer

et al. 2013

Laboratory Investigation

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This paper describes the development of a microfluidic methodology, using RNA extraction and reverse transcription PCR, for investigating expression levels of cytochrome P450 genes. Cytochrome P450 enzymes are involved in the metabolism of xenobiotics, including many commonly prescribed drugs, therefore information on their expression is useful in both pharmaceutical and clinical settings. RNA extraction, from rat liver tissue or primary rat hepatocytes, was performed using a silica-based solid-phase extraction technique. Following elution of the purified RNA, amplification of target sequences for the housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the cytochrome P450 gene CYP1A2, was carried out using a one-step reverse transcription PCR. Once the microfluidic methodology had been optimized, analysis of control and 3-methylcholanthrene-induced primary rat hepatocytes were used to evaluate the system. As expected, GAPDH was consistently expressed, whereas CYP1A2 levels were found to be raised in the drug-treated samples. The proposed system offers an initial platform for development of both rapid throughput analyzers for pharmaceutical drug screening and point-of-care diagnostic tests to aid provision of drug regimens, which can be tailor-made to the individual patient.Laboratory Investigation (2013) 93, 961-966;

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Complexities of Diagnostics of Viruses Affecting Farmed Aquatic Species

Weidmann

2017

Diagnosis and Control of Diseases of Fish and Shellfish

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An integrated microfluidic loop-mediated-isothermal-amplification system for rapid sample pre-treatment and detection of viruses

Cited by 96 publications

References 29 publications

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

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

Integrated RNA extraction and RT-PCR for semi-quantitative gene expression studies on a microfluidic device

Complexities of Diagnostics of Viruses Affecting Farmed Aquatic Species

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