Lee Jeong-Gun, Cheong Kwang Ho, Huh Nam, Kim Suhyeon, Choi Jeong-Woo, Ko Christopher
Bio Lab, Samsung Advanced Institute of Technology, P.O. Box 111, Suwon 440-600, Korea.
Lab Chip. 2006 Jul;6(7):886-95. doi: 10.1039/b515876a. Epub 2006 May 2.
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.
生物样本中病原体的最佳检测取决于快速高效提取DNA分子的能力。在本文中，我们报告了一种通过结合激光照射和磁珠在单个微芯片中进行高效DNA提取及后续实时检测的新方法。通过使用808纳米激光和羧基末端磁珠，我们证明40秒的单脉冲可裂解包括大肠杆菌、革兰氏阳性细菌细胞以及与人类血清混合的乙肝病毒在内的病原体。我们进一步证明，在配备小型激光二极管的手持设备中进行激光照射后，使用同一微芯片在实时PCR仪中用预混PCR试剂进行实时病原体检测。这些结果表明，这种新的样品制备方法非常适合集成到病原体检测系统的芯片实验室应用中。
