A lab-on-chip for biothreat detection using single-molecule DNA mapping.
Rapid, specific, and sensitive detection of airborne bacteria, viruses, and toxins is critical for biodefense, yet the diverse nature of the threats poses a challenge for integrated surveillance, as each class of pathogens typically requires different detection strategies. Here, we present a laboratory-on-a-chip microfluidic device (LOC-DLA) that integrates two unique assays for the detection of airborne pathogens: direct linear analysis (DLA) with unsurpassed specificity for bacterial threats and Digital DNA for toxins and viruses. The LOC-DLA device also prepares samples for analysis, incorporating upstream functions for concentrating and fractionating DNA. Both DLA and Digital DNA assays are single molecule detection technologies, therefore the assay sensitivities depend on the throughput of individual molecules. The microfluidic device and its accompanying operation protocols have been heavily optimized to maximize throughput and minimize the loss of analyzable DNA. We present here the design and operation of the LOC-DLA device, demonstrate multiplex detection of rare bacterial targets in the presence of 100-fold excess complex bacterial mixture, and demonstrate detection of picogram quantities of botulinum toxoid.
US Genomics, 12 Gill Street, Suite 4700, Woburn, MA, USA. rmeltzer@usgenomics.com
Robert H
RH
Meltzer
Jeffrey R
JR
Krogmeier
Lisa W
LW
Kwok
Richard
R
Allen
Bryan
B
Crane
Joshua W
JW
Griffis
Linda
L
Knaian
Nanor
N
Kojanian
Gene
G
Malkin
Michelle K
MK
Nahas
Vyacheslav
V
Papkov
Saad
S
Shaikh
Kedar
K
Vyavahare
Qun
Q
Zhong
Yi
Y
Zhou
Jonathan W
JW
Larson
Rudolf
R
Gilmanshin
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D001427:Bacterial Toxins / Q000032:analysis; D054045:Biological Warfare Agents; D004247:DNA / Q000032:analysis* / Q000276:immunology; D004867:Equipment Design; D023281:Genomics; D007118:Immunoassay; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D011108:Polymers / Q000737:chemistry; D018511:Systems Integration; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification
D001427:Bacterial Toxins; D054045:Biological Warfare Agents; D011108:Polymers; D004247:DNA