ac400412m_si_001.avi (6.57 MB)
Detection of Mycobacterium tuberculosis Using a Capillary-Array Microsystem with Integrated DNA Extraction, Loop-Mediated Isothermal Amplification, and Fluorescence Detection
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posted on 2016-02-19, 09:43 authored by Dayu Liu, Guangtie Liang, Qiong Zhang, Bin ChenThis article reports for the first
time a high-throughput microfluidic system with fully integrated loop-mediated
isothermal amplification (LAMP) analysis. With the developed system,
parallel Mycobacterium tuberculosis detections were
implemented in polytetrafluoroethylene capillaries through the utilization
of droplet technology coupled with magnetic beads. During the analysis,
liquid plugs containing different types of sample or reagents are
sequentially introduced into the capillaries and made to form droplets
therein. The whole analytical process, including DNA extraction, LAMP,
and detection of the amplified products were conducted in such droplets.
The developed microsystem is able to process 10 samples in parallel.
The entire diagnostic procedure, from sample-in to answer-out, can
be automatically completed within 50 min with a limit of detection
(LOD) of 10 bacteria. This microsystem was evaluated by analyzing
clinical samples, and a clinical sensitivity (positive detection rate)
of 96.8% and specificity (negative detection rate) of 100% were achieved.
The presented capillary LAMP assay features high-throughput and low-cost
and thus is a promising tool for rapid tuberculosis diagnosis.