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  • Point-of-care multiplexed assays of nucleic acids using microcapillary-based loop-mediated isothermal amplification.

Point-of-care multiplexed assays of nucleic acids using microcapillary-based loop-mediated isothermal amplification.

Analytical chemistry (2014-06-18)
Yi Zhang, Lu Zhang, Jiashu Sun, Yulei Liu, Xingjie Ma, Shangjin Cui, Liying Ma, Jianzhong Jeff Xi, Xingyu Jiang
ABSTRACT

This report demonstrates a straightforward, robust, multiplexed and point-of-care microcapillary-based loop-mediated isothermal amplification (cLAMP) for assaying nucleic acids. This assay integrates capillaries (glass or plastic) to introduce and house sample/reagents, segments of water droplets to prevent contamination, pocket warmers to provide heat, and a hand-held flashlight for a visual readout of the fluorescent signal. The cLAMP system allows the simultaneous detection of two RNA targets of human immunodeficiency virus (HIV) from multiple plasma samples, and achieves a high sensitivity of two copies of standard plasmid. As few nucleic acid detection methods can be wholly independent of external power supply and equipment, our cLAMP holds great promise for point-of-care applications in resource-poor settings.

MATERIALS
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Sigma-Aldrich
Manganese(II) chloride tetrahydrate, ACS reagent, ≥98%