BackgroundTuberculosis (TB) is a major global health problem and remains the leading cause of morbidity and mortality in developing countries. Routinely used TB diagnostic methods, in most endemic areas, are time-consuming, often less-sensitive, expensive and inaccessible to most patients. Therefore, there is an urgent need for the development of early, easy to use and effective diagnosis tools of TB, which can be effectively integrated into resource limited settings, to anticipate the early treatment and limit further spread of the disease.Over the last decade, Loop-mediated isothermal amplification (LAMP) assays have become a powerful tool for rapid diagnosis of infectious diseases because of the simplicity of device requirements. Indeed, LAMP is a simple, quick and cost effective Isothermal Nucleic Acid Amplification diagnostic test (INAAT) that has the potential to be used in TB endemic settings of resource-poor countries.MethodsIn the present study, we have developed a simple and rapid TB molecular diagnostic test using a Single-Step Loop-mediated isothermal DNA amplification (SS-LAMP) method for the detection of Mycobacterium tuberculosis complex (MTBC) strains, with a simplified sample preparation procedure, eliminating DNA extraction prior to LAMP amplification, DNA initial denaturation and enzymatic inactivation steps during the amplification process.To perform our in-house SS-LAMP assay, a set of six specific primers was specifically designed to recognize eight distinct regions on the MTBC species-specific repetitive insertion sequence 6110 (IS6110). The amplification of the targeted DNA was carried out under isothermal conditions at 65 °C within 1 h. Our protocol was firstly optimized using 60 of confirmed MTBC isolates and a recombinant pGEMeasy-IS6110 vector for sensitivity testing. Thereafter, the assay was evaluated on liquefied sputum specimens collected from 157 Moroccan patients suspected of having TB.ResultsOur SS-LAMP developed assay was able to detect MTBC DNA directly from liquefied sputum samples without any prior DNA extraction, denaturation nor the final enzymatic inactivation step. When compared to routinely used Löwenstein Jensen (LJ) Culture method, our SS-LAMP assay is rapid and showed specificity and sensitivity of 99.14 % and 82.93 % respectively which are within the international standards. In addition, the limit of detection of our assay was found to be as little as 10 copies of bacterial DNA.ConclusionTo our knowledge, this is the first study using a single step LAMP (SS-LAMP) procedure as a rapid, easy to perform and cost effective testing for TB early detection. This innovative assay could be suitable for low-income countries with restricted health equipment facilities.
Wearable sweat sensors offer the possibility of continuous real-time health monitoring of an individual at a low cost without invasion. A variety of sweat glucose sensors have been developed thus far to help diabetes patients frequently monitor blood glucose levels through sweat glucose as a surrogate marker. The present study demonstrates the development and characterization of a three-dimensional paper-based microfluidic electrochemical integrated device (3D PMED) for measuring glucose concentration in sweat in real-time via simple, non-invasive, capillary-action-based sample collection. The device was selective for glucose, and it detected glucose accurately in the clinically relevant range (0~2 mM) in an off-body setup. To the best of our knowledge, this is the first time NEXAR™ has been used for biosensing applications. Further, the developed glucose sensor has acceptable sensitivity of 16.8 µA/mM/cm2. Importantly, in an on-body setup, the device achieved a significant amperometric response to sweat glucose in a very short amount of time (a few seconds). With detailed investigations, this proof-of-concept study could help further the development of sensitive and selective sweat-based glucose sensing devices for real-time glucose monitoring in diabetes patients.
The efficient monitoring of the environment is currently gaining a continuous growing interest in view of finding solutions for the global pollution issues and their associated climate change. In this sense, two-dimensional (2D) materials appear as one of highly attractive routes for the development of efficient sensing devices due, in particular, to the interesting blend of their superlative properties. For instance, graphene (Gr) and graphitic carbon nitride g-C3N4 (g-CN) have specifically attracted great attention in several domains of sensing applications owing to their excellent electronic and physical-chemical properties. Despite the high potential they offer in the development and fabrication of high-performance gas-sensing devices, an exhaustive comparison between Gr and g-CN is not well established yet regarding their electronic properties and their sensing performances such as sensitivity and selectivity. Hence, this work aims at providing a state-of-the-art overview of the latest experimental advances in the fabrication, characterization, development, and implementation of these 2D materials in gas-sensing applications. Then, the reported results are compared to our numerical simulations using density functional theory carried out on the interactions of Gr and g-CN with some selected hazardous gases’ molecules such as NO2, CO2, and HF. Our findings conform with the superior performances of the g-CN regarding HF detection, while both g-CN and Gr show comparable detection performances for the remaining considered gases. This allows suggesting an outlook regarding the future use of these 2D materials as high-performance gas sensors.
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