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首页> 外文会议>Microfluidics, BioMEMS, and Medical Microsystems IV >Rapid and automated sample preparation for nucleic acid extraction on a microfluidic CD (compact disk)
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Rapid and automated sample preparation for nucleic acid extraction on a microfluidic CD (compact disk)

机译：快速自动化的样品制备，可在微流体CD（光盘）上提取核酸

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摘要

Rapid and automated preparation of PCR (polymerase chain reaction)-ready genomic DNA was demonstrated on a multiplexed CD (compact disk) platform by using hard-to-lyse bacterial spores. Cell disruption is carried out while bead-cell suspensions are pushed back and forth in center-tapered lysing chambers by angular oscillation of the disk -keystone effect. During this lysis period, the cell suspensions are securely held within the lysing chambers by heat-activated wax valves. Upon application of a remote heat to the disk in motion, the wax valves release lysate solutions into centrifuge chambers where cell debris are separated by an elevated rotation of the disk. Only debris-free DNA extract is then transferred to collection chambers by capillary-assisted siphon and collected for heating that inactivates PCR inhibitors. Lysing capacity was evaluated using a real-time PCR assay to monitor the efficiency of Bacillus globigii spore lysis. PCR analysis showed that 5 minutes' CD lysis run gave spore lysis efficiency similar to that obtained with a popular commercial DNA extraction kit (i.e., IDI-lysis kit from GeneOhm Sciences Inc.) which is highly efficient for microbial cell and spore lysis. This work will contribute to the development of an integrated CD-based assay for rapid diagnosis of infectious diseases.
机译：通过使用难裂解的细菌孢子，在多重CD（光盘）平台上证明了快速，自动化的PCR（聚合酶链反应）就绪基因组DNA的制备方法。通过圆盘-梯形失真效应的角振荡，在中心锥形的裂解室中前后推动珠状细胞悬浮液的同时，进行细胞破坏。在此裂解期间，细胞悬浮液通过热激活蜡阀牢固地保持在裂解室内。在对运动的圆盘施加少量热量后，蜡阀将裂解液释放到离心室中，在那里通过圆盘的旋转旋转将细胞碎片分离。然后，只有无碎片的DNA提取物通过毛细管辅助虹吸管转移到收集室，并收集加热以灭活PCR抑制剂。使用实时PCR测定来评估裂解能力以监测球形芽孢杆菌孢子裂解的效率。 PCR分析表明5分钟的CD裂解运行产生的孢子裂解效率类似于使用流行的商业DNA提取试剂盒（即，GeneOhm Sciences Inc.的IDI裂解试剂盒）获得的，其对于微生物细胞和孢子裂解非常有效。这项工作将有助于开发基于CD的集成化分析方法，以快速诊断传染病。

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