Multiplexed detection and differentiation of the DNA strains for influenza A (H1N1 2009) using a silicon-based microfluidic system
| Autor: | Guang Kai Ignatius Tay, Lakshmi Shankar, Linus Tzu-Hsiang Kao, Tae Goo Kang, Siti Rafeah Mohamed Rafei, Guo-Jun Zhang, Charlie Wah Heng Lee |
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| Rok vydání: | 2010 |
| Předmět: |
DNA
Bacterial Silicon Microfluidics Biomedical Engineering Biophysics Computational biology Biosensing Techniques Biology medicine.disease_cause Multiplexing Polymerase Chain Reaction chemistry.chemical_compound Influenza A Virus H1N1 Subtype Electrochemistry Influenza A virus medicine Silicon nanowires Point of care Oligonucleotide Array Sequence Analysis General Medicine Equipment Design Sequence Analysis DNA Amplicon Microfluidic Analytical Techniques Virology Equipment Failure Analysis chemistry Nucleic acid DNA Biotechnology |
| Zdroj: | Biosensorsbioelectronics. 26(5) |
| ISSN: | 1873-4235 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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