Search

Complexity of sample preparation decelerate the development of sample-in-answer-out devices for point-of-need nucleic acid amplification testing. Here, we present the consolidation of alkaline poly(ethylene) glycol-based lysis and solid-phase extraction for rapid and simple sample preparation compatible with direct on-bead amplification. Simultaneous cell lysis and binding of DNA were achieved using an optimised reagent comprising 15% PEG8000, 0.5 M NaCl, and 3.5 mM KOH. This was combined with direct, on-bead amplification using 1.5 μg beads per 20 μL PCR reaction mix. The novel single reagent, 5-min method improved the detection limit by 10 and 100-fold compared with commercial DNA extraction kits and the original alkaline PEG lysis method, respectively. The sensitivity can be further enhanced by one amplification cycle with an ethanol wash or by extending the incubation to 10 min before collecting the magnetic particles. Both methods successfully detected a single copy of Escherichia coli DNA. In biological fluids (saliva, sweat, and urine), the 5-min method was delayed by about one cycle compared to the 15-min method. The proposed methods are attractive for incorporation in the workflow for point-of-need testing of biological samples by providing a practical and chemical method for simple alternative DNA sample preparation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Nucleic acid amplification testing facilitates the detection of disease through specific genomic sequences and is attractive for point-of-need testing (PONT); in particular, the early detection of microorganisms can alert early response systems to protect the public and ecosystems from widespread outbreaks of biological threats, including infectious diseases. Prior to nucleic acid amplification and detection, extensive sample preparation techniques are required to free nucleic acids and extract them from the sample matrix. Sample preparation is critical to maximize the sensitivity and reliability of testing. As the enzymatic amplification reactions can be sensitive to inhibitors from the sample, as well as from chemicals used for lysis and extraction, avoiding inhibition is a significant challenge, particularly when minimising liquid handling steps is also desirable for the translation of the assay to a portable format for PONT. The reagents used in sample preparation for nucleic acid testing, covering lysis and NA extraction (binding, washing, and elution), are reviewed with a focus on their suitability for use in PONT.

Introduction: Ethambutol is an antibiotic used as a first line drug in the treatment of tuberculosis and a vision threatening side effect of EMB is ethambutol-induced optic neuropathy (EON). The aim of the study is to create awareness about the potentiality of ethambutol to cause ethambutol-induced optic neuropathy, careful monitoring of dose and patient education., Materials and Methods: A retrospective observational study of 14 patients whose complete Anti- tubercular treatment records could be retrieved were included. Epidemiological data including age, sex, systemic illness were recorded. Duration between optic nerve toxicity , usage of ethambutol and the drug dosage were noted. Best corrected visual acuity, anterior segment examination including pupils, extraocular movements, colour vision, central fields and fundus examination were evaluated. The patients were followed up at one and three month intervals., Results: Associated systemic illness was found to be a confounding factor for the development of ethambutol-induced optic neuropathy. 57% of patients had diabetes mellitus followed by hypertension (14.2%), renal disease (7.1%). The average daily dose of Ethambutol ingested was 1078.5 mg (21 mg/kg) and this high dose could have been the primary cause for development of ethambutol-induced optic neuropathy. Vision ranged from total blindness to mild visual impairment and poor recovery of vision was noted even after discontinuing ethambutol., Conclusion: Only a minority of patients showed improvement in visual function following discontinuation of ethambutol and the toxicity was found to be dose-dependent. Patients with comorbidities like renal impairment and diabetes mellitus appeared to be at greater risk. Ophthalmological examination before commencing treatment and periodic evaluation thereafter is mandatory., (© NEPjOPH.)

Porous materials facilitate the efficient separation of chemicals and particulate matter by providing selectivity through structural and surface properties and are attractive as sorbent owing to their large surface area. This broad applicability of porous materials makes the integration of porous materials and microfluidic devices important in the development of more efficient, advanced separation platforms. Additive manufacturing approaches are fundamentally different to traditional manufacturing methods, providing unique opportunities in the fabrication of fluidic devices. The complementary 3D printing (3DP) methods are each accompanied by unique opportunities and limitations in terms of minimum channel size, scalability, functional integration and automation. This review focuses on the developments in the fabrication of 3DP miniaturised fluidic devices with integrated porous materials, focusing polymer-based methods including fused filament fabrication (FFF), inkjet 3D printing and digital light projection (DLP). The 3DP methods are compared based on resolution, scope for multimaterial printing and scalability for manufacturing. As opportunities for printing pores are limited by resolution, the focus is on approaches to incorporate materials with sub-micron pores to be used as membrane, sorbent or stationary phase in separation science using Post-Print, Print-Pause-Print and In-Print processes. Technical aspects analysing the efficiency of the fabrication process towards scalable manufacturing are combined with application aspects evaluating the separation and/or extraction performance. The review is concluded with an overview on achievements and opportunities for manufacturable 3D printed membrane/sorbent integrated fluidic devices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

In the past decade, 3D printing technologies have been adopted for the fabrication of microfluidic devices. Extrusion-based approaches including fused filament fabrication (FFF), jetting technologies including inkjet 3D printing, and vat photopolymerization techniques including stereolithography (SLA) and digital light projection (DLP) are the 3D printing methods most frequently adopted by the microfluidic community. Each printing technique has merits toward the fabrication of microfluidic devices. Inkjet printing offers a good selection of materials and multimaterial printing, and the large build space provides manufacturing throughput, while FFF offers a great selection of materials and multimaterial printing but at lower throughput compared to inkjet 3D printing. Technical and material developments adopted from adjacent research fields and developed by the microfluidic community underpin the printing of sub-100 μm enclosed microchannels by DLP, but challenges remain in multimaterial printing throughput. With the feasibility of 3D printed microfluidics established, we look ahead at trends in 3D printing to gain insights toward the future of this technology beyond the sole prism of being an alternative fabrication approach. A shift in emphasis from using 3D printing for prototyping, to mimic conventionally manufactured outputs, toward integrated approaches from a design perspective is critically developed.