Multiplexed detection and differentiation of the DNA strains for influenza A (H1N1 2009) using a silicon-based microfluidic system

• Published in: Biosensors & bioelectronics 2011-01, Vol.26 (5), p.2006-2011
• Main Authors: Kao, Linus Tzu-Hsiang, Shankar, Lakshmi, Kang, Tae Goo, Zhang, Guojun, Tay, Guang Kai Ignatius, Rafei, Siti Rafeah Mohamed, Lee, Charlie Wah Heng
• Format: Article
• Language: English
• Subjects: Airports; Biological and medical sciences; Biosensing Techniques - instrumentation; Biosensors; Biotechnology; Differentiation; Disease control; DNA, Bacterial - genetics; DNA, Bacterial - isolation & purification; Equipment Design; Equipment Failure Analysis; Fundamental and applied biological sciences. Psychology; Infectious diseases; Influenza A; Influenza A Virus, H1N1 Subtype - classification; Influenza A Virus, H1N1 Subtype - genetics; Influenza A Virus, H1N1 Subtype - isolation & purification; Influenza virus; Microfluidic Analytical Techniques - instrumentation; Microfluidic PCR; Microfluidics; nanotechnology; nucleic acids; Oligonucleotide Array Sequence Analysis - instrumentation; pandemics; Point-of-care diagnosis; Polymerase chain reaction; Polymerase Chain Reaction - instrumentation; Sequence Analysis, DNA - instrumentation; Silicon; Silicon - chemistry; Silicon nanowire; Swine flu (H1N1 2009)
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2010.08.076

Pandemic influenza by the swine-origin influenza virus (H1N1 2009) has attracted considerable concern worldwide. A convenient and accurate diagnostic approach that can be deployed at the point of care, such as in a doctor's office or at an airport, is critical for disease control. Here we report the development of a silicon-based microfluidic system for subtype differentiation of the novel H1N1 2009 strain vs. the seasonal influenza A (FluA) strain. The proposed system included two functional modules: a multiplexed PCR module for amplification of nucleic acid targets and a multiplexed silicon nanowire (SiNW) module for sequence determination. The PCR module consisted of a microfluidic PCR chamber and an electrical controller to perform a multiplexed protocol that simultaneously enriched specific segments of both H1N1 and FluA strains (if present), with 10 4–10 5 amplification efficiency. The PCR amplicon was subsequently denatured and transferred to the SiNW sensing module for a label-free, multiplexed detection. A control SiNW was implemented, for the first time, in order to eliminate background interference. The detection module demonstrated a 10× change in the magnitude of differential current when the target DNA was injected. Overall, the system achieved a sensitivity of 20–30 fg/μl for H1N1 and seasonal FluA nucleic acids in a 10 μl sample. The low sample consumption, high sensitivity and high specificity render it a potential point-of-care (POC) platform to help doctors reach a yes/no decision for infectious diseases.