Loop-mediated isothermal amplification in disposable polyester-toner microdevices

Oliveira, Kézia Gomes de; Borba, Juliane C.; Bailão, Alexandre Melo; Soares, Célia Maria de Almeida; Carrilho, Emanuel; Duarte, Gabriela Rodrigues Mendes

doi:10.1016/j.ab.2017.07.014

Cited by 6 publications

(9 citation statements)

References 57 publications

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“…The best results obtained previously required passivation with BSA (0.6 mg mL −1 ). In our previous study, 41 we also found the importance of controlling the A / V (area / volume) to improve the efficiency of the amplification reaction. The results showed that the best A / V ratio was 6 mm 2 μL −1 (channel dimensions: 7 mm in length, 2 mm in width and 0.35 mm in depth).…”

Section: Resultsmentioning

confidence: 82%

“…We previously demonstrated that PeT devices could be successfully used for DNA amplification by LAMP. 41 In our first paper about isothermal DNA amplification in a PeT microchip, we demonstrated E. coli detection with high sensitivity. We also confirmed that it is necessary to passivate the inner surfaces of the channels to avoid the absorption of reagents on microchamber.…”

Section: Resultsmentioning

confidence: 95%

“…Before use, the microchambers were passivated with BSA (1.0 mg mL −1 ), as previously described. 41 After passivation, the LAMP chamber was filled with approximately 5 µL of master mix solution by capillary force. Mineral oil overlaid on both reservoirs prevented evaporation of the solution.…”

Section: Rt-lamp Amplification Of Dengue Virus In Pet Microchipmentioning

confidence: 99%

“…[37][38][39] Polyester-toner (PeT) microchips have been widely used in recent years for applications with biological samples, thus demonstrating compatibility for clinical diagnostic applications. [40][41][42][43][44][45][46][47][48][49][50] Both polyester and toner are inexpensive materials and the microfabrication process of the device is simple, fast, and low-cost, in addition to being disposable. 41 In this paper, we describe a simple and rapid method for molecular diagnostics of dengue fever by RT-LAMP in a PeT microchip.…”

Section: Introductionmentioning

confidence: 99%

“…[40][41][42][43][44][45][46][47][48][49][50] Both polyester and toner are inexpensive materials and the microfabrication process of the device is simple, fast, and low-cost, in addition to being disposable. 41 In this paper, we describe a simple and rapid method for molecular diagnostics of dengue fever by RT-LAMP in a PeT microchip. For a proof-of-concept, we detected the DENV-4 in serum samples of infected patients.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Diagnostics of Dengue by Reverse Transcription-Loop Mediated Isothermal Amplification (RT-LAMP) in Disposable Polyester-Toner Microdevices

Mendes¹,

Oliveira²,

Borba³

et al. 2019

J. Braz. Chem. Soc.

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Dengue is one of the most prevalent infectious tropical diseases in the world, with high incidence in over 100 countries. Rapid and reliable diagnosis of dengue is of great importance for public health. To simplify molecular diagnostics, isothermal amplification techniques have recently emerged as an alternative to conventional methods of deoxyribonucleic acid (DNA) amplification. Here, we developed a one-step method for dengue virus detection from real sample based on RT-LAMP (reverse transcription-loop mediated isothermal amplification) in a disposable microdevice. The reaction was thermally controlled with a thermoblock for 15 min at 72 °C. At the end of the incubation time, we either removed the solution for detection of fragments by gel electrophoresis or added DNA intercalator for visual detection on-chip. Our results demonstrated that it is possible to detect dengue virus through RT-LAMP directly from a serum sample, without previous ribonucleic acid (RNA) extraction. The success of RT-LAMP was confirmed in reactions initiated with 0.8 fg μL −1 of RNA, which represents 200 copies of RNA per μL. RT-LAMP in a polyester-toner (PeT) microdevice is a simple and inexpensive method that allows for rapid detection of dengue virus with high reliability and great potential for point-of-care applications.

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Section: Resultsmentioning

confidence: 82%

Section: Resultsmentioning

confidence: 95%

Section: Rt-lamp Amplification Of Dengue Virus In Pet Microchipmentioning

confidence: 99%