A large-volume sputum dry storage and transportation device for molecular and culture-based diagnosis of tuberculosis

Dsouza, Andrea; Jangam, Saylee; Soni, Shruti; Agarwal, Priyanka; Naik, V. S.; Manjula, J; Nair, Chandrasekhar Bhaskaran; Toley, Bhushan J.

doi:10.1039/d1lc00900a

Cited by 2 publications

(3 citation statements)

References 24 publications

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“…For this, a minimum infrastructure should be available when working with this biological medium. It has recently been reported that the collected sputum samples could also be stored and dried at ambient conditions for up to 10 days using a technology developed by A. Dsouza et al, thus facilitating sputum transportation if necessary . Nevertheless, sample processing has to be carried out upon sputum reconstitution and before carrying out the analyses, too.…”

Section: Discussionmentioning

confidence: 99%

“…It has recently been reported that the collected sputum samples could also be stored and dried at ambient conditions for up to 10 days using a technology developed by A. Dsouza et al, thus facilitating sputum transportation if necessary. 53 Nevertheless, sample processing has to be carried out upon sputum reconstitution and before carrying out the analyses, too. The device quantitatively detected the three target biomarkers at those concentrations that appeared to be clinically relevant.…”

Section: Discussionmentioning

confidence: 99%

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Engineering a Point-of-Care Paper-Microfluidic Electrochemical Device Applied to the Multiplexed Quantitative Detection of Biomarkers in Sputum

et al. 2023

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Health initiatives worldwide demand affordable point-of-care devices to aid in the reduction of morbidity and mortality rates of high-incidence infectious and noncommunicable diseases. However, the production of robust and reliable easy-to-use diagnostic platforms showing the ability to quantitatively measure several biomarkers in physiological fluids and that could in turn be decentralized to reach any relevant environment remains a challenge. Here, we show the particular combination of paper-microfluidic technology, electrochemical transduction, and magnetic nanoparticle-based immunoassay approaches to produce a unique, compact, and easily deployable multiplex device to simultaneously measure interleukin-8, tumor necrosis factor-α, and myeloperoxidase biomarkers in sputum, developed with the aim of facilitating the timely detection of acute exacerbations of chronic obstructive pulmonary disease. The device incorporates an on-chip electrochemical cell array and a multichannel paper component, engineered to be easily aligned into a polymeric cartridge and exchanged if necessary. Calibration curves at clinically relevant biomarker concentration ranges are produced in buffer and artificial sputum. The analysis of sputum samples of healthy individuals and acutely exacerbated patients produces statistically significant biomarker concentration differences between the two studied groups. The device can be mass-produced at a low cost, being an easily adaptable platform for measuring other disease-related target biomarkers.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Engineering a Point-of-Care Paper-Microfluidic Electrochemical Device Applied to the Multiplexed Quantitative Detection of Biomarkers in Sputum

et al. 2023

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“…The qPCR cycle was as follows: initial denaturation at 95 °C for 5 minutes followed by 35 cycles of denaturation at 95 °C for 30 s, annealing at 58 °C for 30 s, and extension at 72 °C for 20 s. Fluorescence readings were acquired during the extension step. Mycobacterium smegmatis: Msm bacteria were quantified using a qPCR assay adapted from Hümpel et al 49 and Dsouza et al 50 Primers for the sigA gene of Msm werereverse:…”

Section: Qpcr Assaysmentioning

confidence: 99%

Integrated bacterial cell lysis and DNA extraction using paper-based isotachophoresis – towards a versatile sample preparation module for point-of-care diagnostics

Soni

Toley

2023

Preprint

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Bacterial infections remain a global threat, particularly in low-resource settings, where access to accurate and timely diagnosis is limited. Point-of-care nucleic acid amplification tests have shown great promise in addressing this challenge. However, their dependence on complex traditional sample preparation methods remains a major challenge. To address this limitation, we present a paper-based sample preparation module that integrates bacterial cell lysis, DNA purification, and concentration using an electrokinetic technique called isotachophoresis (ITP). This is the first device that i) integrates electrochemical bacterial lysis with ITP, and ii) demonstrates the focusing of whole bacterial genomic DNA (gDNA) in paper. Characterization with buffers showed that the paper-based ITP sample preparation module (p-ITPrep) concentrated bacterial gDNA with an average concentration factor of 12X, and DNA could be extracted from a sample containing as few as 10^2 CFU/mL Mycobacterium smegmatis (Msm). From complex biological matrices – human saliva, human blood serum, and artificial urine, p-ITPrep extracted DNA from samples containing 10^2 CFU Msm/mL saliva or artificial urine and 10^3 CFU Msm/mL serum within 20 minutes. The extraction procedure involved only 3 user steps, in contrast to conventional solid phase extraction kits that require more than 10 user steps. p-ITPrep may provide a simple, inexpensive, and versatile alternative to conventional multi-step nucleic acid extraction protocols for point-of-care diagnostic.

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A large-volume sputum dry storage and transportation device for molecular and culture-based diagnosis of tuberculosis

Abstract: We present SPECTRA-tube (specimen transportation tube), a device that enables the dry stabilization and transportation of large volumes (>1 mL) of liquid specimens. We demonstrate its application in molecular and culture testing of tuberculosis.

Cited by 2 publications

References 24 publications

Engineering a Point-of-Care Paper-Microfluidic Electrochemical Device Applied to the Multiplexed Quantitative Detection of Biomarkers in Sputum

Engineering a Point-of-Care Paper-Microfluidic Electrochemical Device Applied to the Multiplexed Quantitative Detection of Biomarkers in Sputum

Integrated bacterial cell lysis and DNA extraction using paper-based isotachophoresis – towards a versatile sample preparation module for point-of-care diagnostics

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