Simple system for isothermal DNA amplification coupled to lateral flow detection.
Infectious disease diagnosis in point-of-care settings can be greatly improved through integrated, automated nucleic acid testing devices. We have developed an early prototype for a low-cost system which executes isothermal DNA amplification coupled to nucleic acid lateral flow (NALF) detection in a mesofluidic cartridge attached to a portable instrument. Fluid handling inside the cartridge is facilitated through one-way passive valves, flexible pouches, and electrolysis-driven pumps, which promotes a compact and inexpensive instrument design. The closed-system disposable prevents workspace amplicon contamination. The cartridge design is based on standard scalable manufacturing techniques such as injection molding. Nucleic acid amplification occurs in a two-layer pouch that enables efficient heat transfer. We have demonstrated as proof of principle the amplification and detection of Mycobacterium tuberculosis (M.tb) genomic DNA in the cartridge, using either Loop Mediated Amplification (LAMP) or the Exponential Amplification Reaction (EXPAR), both coupled to NALF detection. We envision that a refined version of this cartridge, including upstream sample preparation coupled to amplification and detection, will enable fully-automated sample-in to answer-out infectious disease diagnosis in primary care settings of low-resource countries with high disease burden.
Keck Graduate Institute of Applied Life Sciences, Claremont, California, United States of America.
Kristina
K
Roskos
Anna I
AI
Hickerson
Hsiang-Wei
HW
Lu
Tanya M
TM
Ferguson
Deepali N
DN
Shinde
Yvonne
Y
Klaue
Angelika
A
Niemz
D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification*; D044085:Microfluidics; D009169:Mycobacterium tuberculosis / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D012212:Rheology / Q000295:instrumentation / Q000379:methods*; D013696:Temperature
D004269:DNA, Bacterial