(101e) Multiplexed Detection of Viral Nucleic Acids In a Combinatorial Microfluidic Screening Chip

In this work, we report a multiplexed microfluidic device for detection of nucleic acids for health care applications. Accurate medical diagnosis often requires multiple clinical tests measuring and/or screening for a number of health or disease indicators. At present, however, most diagnostic tests only provide information on a single indicator or marker. For this reason, rapid multiplexed assays based on integrated microdevices could significantly improve the quality of healthcare by reducing turnaround time and analysis cost, and speeding up the decision making process for diagnosis and treatment. Specifically, microfluidic techniques offer several key advantages for diagnostic applications, including miniaturized instrumentation, reduced amounts of sample and reagents for analysis, and integration of assay steps such as sample preparation (processing of physiological fluids), post-processing (e.g. reagent mixing, labeling, separation), detection and analysis into a single device.

Here, we describe a combinatorial screening chip for high sensitivity, sequence-specific screening and detection of viral nucleic acid markers (ssDNA) using surface immobilized molecular beacons and total internal reflection fluorescence microscopy (TIRFM). Using a 4 x 4 array of 200 pL wells, we screened for the presence of four target single stranded oligonucleotides encoding for conserved regions of the genomes of four common viruses: human immunodeficiency virus-1 (HIV-1), human papillomavirus (HPV), Hepatitis A (Hep A) and Hepatitis B (Hep B). Target oligonucleotides are accurately detected and discriminated against alternative oligonucleotides with different sequences. The combinatorial chip represents a versatile platform for the development of clinical diagnostic tests for simultaneous screening, detection and monitoring of a wide range of biological markers of health and disease using minimal sample size.

1.   B. R. Schudel, M. Tanyeri, A. Mukherjee, C. M. Schroeder and Paul J. A. Kenis, Lab on a Chip, 11, (2011).