Thermal lysis and isothermal amplification of Mycobacterium tuberculosis H37Rv in one tube

Shetty, Prasad; Ghosh, Debanjan; Paul, Debjani

doi:10.1016/j.mimet.2017.09.013

Cited by 16 publications

(10 citation statements)

References 14 publications

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“…The plant DNA extraction process used in the above experiments is not very simple or user-friendly, as it requires repeated centrifugation steps and careful removal of certain supernatants. Several groups have worked to develop simpler nucleic acid extraction procedures [21][22][23] that enable an untrained user to perform the complete assay. Our lab is also currently developing methods to simplify the extraction procedure.…”

Section: Discussionmentioning

confidence: 99%

Distance and Microsphere Aggregation-Based DNA Detection in a Paper-Based Microfluidic Device

et al. 2020

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In paper-based microfluidics, the simplest devices are colorimetric, giving qualitative results. However, getting quantitative data can be quite a bit more difficult. Distance-based devices provide a user-friendly means of obtaining quantitative data without the need for any additional equipment, simply by using an included ruler or calibrated markings. This article details the development of a quantitative DNA detection device that utilizes the aggregation of polystyrene microspheres to affect the distance that microspheres wick through filter paper. The microspheres are conjugated to single-stranded DNA (ssDNA) oligomers that are partially complementary to a target strand and, in the presence of the target strand, form a three-strand complex, resulting in the formation of aggregates. The higher the concentration of the target strand, the larger the aggregate, and the shorter the distance wicked by the microspheres. This behavior was investigated across a wide range of target concentrations and under different incubation times to understand aggregate formation. The device was then used to successfully detect a target strand spiked in extracted plant DNA.

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Section: Discussionmentioning

confidence: 99%

Distance and Microsphere Aggregation-Based DNA Detection in a Paper-Based Microfluidic Device

et al. 2020

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“…Thermal lysis of cells is conducted by exposing them to an elevated temperature of 80 � C or more (Shetty et al, 2017) to rupture their membranes and release the NAs. This method can be used to direct the release of DNA from the spores (P� ribylka et al, 2013), which is normally a difficult or time consuming process.…”

Section: On-chip Cell Lysismentioning

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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“…Our aim was to integrate the sample preparation and DNA amplification steps into a single reaction step without any intermediate DNA purification. We demonstrated an integrated one-tube workflow at 65 °C and 60 min that completely disinfected the MTB (H37Rv) culture, thermally lysed the bacteria and amplified the DNA by HDA (Shetty et al 2017) (Fig. 2).…”

Section: Our Past Work In Molecular Diagnosticsmentioning

confidence: 99%

“…b Thermal disinfection of pathogenic bacte-ria, lysis and HDA achieved in a single heat incubation step at 65 °C and 60 min (lanes 9, 10). Reproduced fromShetty et al (2017) with permission from Elsevier…”

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confidence: 99%

Developing a Point-of-Care Molecular Test to Detect SARS-CoV-2

Paul

Naik

Roy

2020

Trans Indian Natl. Acad. Eng.

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There is a need for widespread testing in India to stop the spread of the novel coronavirus in the population. While RT-PCR is the recommended diagnostic technique, its use is limited to well-equipped laboratories due to the need for specialized instrumentation, reagents and trained personnel. Immunodiagnostic tests are not yet recommended by the WHO for diagnosing active infections. There is a strong need for developing point-of-care molecular tests. Based on our past experience with paperfluidic devices for diagnosing bacterial infections by molecular tests, we propose the development of a diagnostic test for COVID-19. As a platform technology, it could be adapted to other viral outbreaks in future.

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Thermal lysis and isothermal amplification of Mycobacterium tuberculosis H37Rv in one tube

Cited by 16 publications

References 14 publications

Distance and Microsphere Aggregation-Based DNA Detection in a Paper-Based Microfluidic Device

Distance and Microsphere Aggregation-Based DNA Detection in a Paper-Based Microfluidic Device

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

Developing a Point-of-Care Molecular Test to Detect SARS-CoV-2

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