Microchip-based one step DNA extraction and real-time PCR in one chamber for rapid pathogen identification.
Optimal detection of a pathogen present in biological samples depends on the ability to extract DNA molecules rapidly and efficiently. In this paper, we report a novel method for efficient DNA extraction and subsequent real-time detection in a single microchip by combining laser irradiation and magnetic beads. By using a 808 nm laser and carboxyl-terminated magnetic beads, we demonstrate that a single pulse of 40 seconds lysed pathogens including E. coli and Gram-positive bacterial cells as well as the hepatitis B virus mixed with human serum. We further demonstrate that the real-time pathogen detection was performed with pre-mixed PCR reagents in a real-time PCR machine using the same microchip, after laser irradiation in a hand-held device equipped with a small laser diode. These results suggest that the new sample preparation method is well suited to be integrated into lab-on-a-chip application of the pathogen detection system.
Lee
Jeong-Gun
JG
Bio Lab, Samsung Advanced Institute of Technology, P.O. Box 111, Suwon 440-600, Korea. biogun.lee@samsung.com
Cheong
Kwang Ho
KH
Huh
Nam
N
Kim
Suhyeon
S
Choi
Jeong-Woo
JW
Ko
Christopher
C
D004269:DNA, Bacterial / Q000302:isolation & purification*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D013212:Staphylococcus epidermidis / Q000737:chemistry / Q000235:genetics; D013295:Streptococcus mutans / Q000737:chemistry / Q000235:genetics
D004269:DNA, Bacterial