An integrated, stacked microlaboratory for biological agent detection with DNA and immunoassays

• Published in: Biosensors & bioelectronics 2002-06, Vol.17 (6), p.605-618
• Main Authors: Yang, Joon Mo, Bell, Janice, Huang, Ying, Tirado, Marcus, Thomas, Donald, Forster, Anita H, Haigis, Robert W, Swanson, Paul D, Wallace, R.Bruce, Martinsons, Bob, Krihak, Michael
• Format: Article
• Language: English
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Biological Warfare - prevention & control; Biosensing Techniques - instrumentation; Biosensing Techniques - methods; Biotechnology; DNA hybridization assay; DNA Probes - chemistry; DNA, Bacterial - analysis; DNA, Bacterial - genetics; Electrochemistry - methods; Electromagnetic Fields; Electrophoresis - methods; Enterotoxins - analysis; Equipment Design; Escherichia coli - classification; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Immunoassay; Immunoassay - instrumentation; Immunoassay - methods; Integrated fluidic system; Integrated protocol; Methods. Procedures. Technologies; Microbiology; Miniaturization; Nucleic Acid Amplification Techniques - instrumentation; Nucleic Acid Amplification Techniques - methods; Nucleic Acid Hybridization; Others; SDA module; Semiconductors; Shiga Toxin 1 - analysis; Shiga Toxin 1 - genetics; Stacked structure; Various methods and equipments
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/S0956-5663(02)00023-4

An integrated, stacked microlaboratory for performing automated electric-field-driven immunoassays and DNA hybridization assays was developed. The stacked microlaboratory was fabricated by orderly laminating several different functional layers (all 76×76 mm 2) including a patterned polyimide layer with a flip-chip bonded CMOS chip, a pressure sensitive acrylic adhesive (PSA) layer with a fluidic cutout, an optically transparent polymethyl methacrylate (PMMA) film, a PSA layer with a via, a patterned polyimide layer with a flip-chip bonded silicon chip, a PSA layer with a fluidic cutout, and a glass cover plate layer. Versatility of the stacked microlaboratory was demonstrated by various automated assays. Escherichia coli bacteria and Alexa-labeled protein toxin staphylococcal enterotoxin B (SEB) were detected by electric-field-driven immunoassays on a single chip with a specific-to-nonspecific signal ratios of 4.2:1 and 3.0:1, respectively. Furthermore, by integrating the microlaboratory with a module for strand displacement amplification (SDA), the identification of the Shiga-like toxin gene (SLT1) from E. coli was accomplished within 2.5 h starting from a dielectrophoretic concentration of intact E. coli bacteria and finishing with an electric-field-driven DNA hybridization assay, detected by fluorescently labeled DNA reporter probes. The integrated microlaboratory can be potentially used in a wide range of applications including detection of bacteria and biowarfare agents, and genetic identification.