Development of an Automated Microfluidic System for DNA Collection, Amplification, and Detection of Pathogens

Hagan, Bethany S. ; Bruckner-Lea, Cynthia J.

Pacific Northwest National Laboratory (U.S.)

关键词: Dna;    Pcr;    Sda;    Testing Pathogen Detection;    Polymerase Chain Reaction;   

DOI  :  10.2172/974521
RP-ID  :  PNNL-13951
RP-ID  :  AC05-76RL01830
RP-ID  :  974521

美国|英语

来源: UNT Digital Library

【 摘 要 】

This project was focused on developing and testing automated routines for a microfluidic Pathogen Detection System. The basic pathogen detection routine has three primary components; cell concentration, DNA amplification, and detection. In cell concentration, magnetic beads are held in a flow cell by an electromagnet. Sample liquid is passed through the flow cell and bacterial cells attach to the beads. These beads are then released into a small volume of fluid and delivered to the peltier device for cell lysis and DNA amplification. The cells are lysed during initial heating in the peltier device, and the released DNA is amplified using polymerase chain reaction (PCR) or strand displacement amplification (SDA). Once amplified, the DNA is then delivered to a laser induced fluorescence detection unit in which the sample is detected. These three components create a flexible platform that can be used for pathogen detection in liquid and sediment samples. Future developments of the system will include on-line DNA detection during DNA amplification and improved capture and release methods for the magnetic beads during cell concentration.