Single-use, electricity-free amplification device for detection of HIV-1

Abstract

Early and accurate diagnosis of HIV is key for the reduction of transmission and initiation of patient care. The availability of a rapid nucleic acid test (NAT) for use at the point-of-care (POC) will fill a gap in HIV diagnostics, improving the diagnosis of acute infection and HIV in infants born to infected mothers. In this study, we evaluated the performance of non-instrumented nucleic acid amplification, single-use disposable (NINA-SUD) devices for the detection of HIV-1 in whole blood using reverse-transcription, loop-mediated isothermal amplification (RT-LAMP) with lyophilized reagents. The NINA-SUD heating device harnesses the heat from an exothermic chemical reaction initiated by the addition of saline to magnesium iron powder. Reproducibility was demonstrated between NINA-SUD units and comparable, if not superior, performance for detecting clinical specimens was observed as compared to the thermal cycler. The stability of the lyophilized HIV-1 RT-LAMP reagents was also demonstrated following storage at −20, 4, 25, and 30°C for up to one month. The single-use, disposable NAT minimizes hands-on time and has the potential to facilitate HIV-1 testing in resource-limited settings or at the POC.

Evaluation of the stability of lyophilized loop-mediated isothermal amplification reagents for the detection of Coxiella burnetii

“…A number of lyophilized LAMP and RT-LAMP assays have been developed for detection of bacteria, virus and parasites [ 33 , 34 , 35 , 36 , 37 ]. Carter et al showed that the lyophilized reagent mixture remained stable at room temperature with little loss of activity for over one and half months [36] .…”

Loop-mediated isothermal amplification (LAMP) – review and classification of methods for sequence-specific detection

“…The insertion of biotin into amplicons by biotin-labelled primers is unproblematic regarding the efficiency of amplication due to Aedes aegypti (Wolbachia infection) 85,86 Ammonia-oxidizing enzyme in environmental bacteria 64 Avian reovirus 70 Bacteriophage MS2 73 Bemisia tabaci 60 BRAF allele (V600E) 83 Brucella 72 Caenorhabditis elegans 68 Chikungunya virus 73 Chlamydia trachomatis 61 Dengue virus 87,112,156 Diarrheal disease 82 Escherichia coli 68 Fomitiporia torreyae 59 Fulviformes umbrinellus 59 Fusarium oxysporum f. sp. lycopersici (point mutations in xylem 3 (SIX3) gene) 66 Genetically modied maize [77][78][79][80][81] Haemophilus ducreyi 101 Haemophilus inuenzae 105 hBRCA1 68 HeLa 68 Hepatitis B and C virus 62,87,107 Herpes simplex virus 1 (HSV1) US4 83 Human immunodeciency virus 87,101 HLA-B*15:02 allele 163 Human papillomavirus 153,171 Human T-lymphotropic virus 1 101 Inuenza virus 63,114 Lambda DNA 68 Leptospira 148 Leishmania 155 Listeria ivanovii 103 Listeria monocytogenes 103 Methicillin-resistant Staphylococcus aureus (MRSA) 71,166 Middle east respiratory sy...…”

“…Similar approaches allowing visual endpoint detection through the addition of quencher oligonucleotides aer amplication were demonstrated for the human immunodeciency virus (HIV) and genetically modied maize. [77][78][79][80][81] Detection of diarrheal disease by QUASR was performed on a centrifugal microuidic platform. 82 QUASR is limited to biplex LAMP if visual detection is performed due to overlapping of the colours.…”

Single-use paper-based hydrogen fuel cells for point-of-care diagnostic applications

“…The duration of the power delivery can be extended as these devices use up to 20x more MgFe for a single test (and consequently, produce as much hydrogen) than the amount used in the fuel cells presented here. [39][40][41][42] This approach implies reusability of the fuel cells for more than one measurement, thus reducing the overall cost per test.…”

“…the corrosion of magnesium iron particles. [36][37][38][39][40][41][42] This chemistry is very convenient, as it , requires only the addition of water to achieve 100°C (a reason why it has also been widely used to heat portable meals). During the course of the chemical reaction the magnesium is oxidized, releasing hydrogen gas as a byproduct, which previously was simply vented off to the atmosphere.…”