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A suction-type microfluidic immunosensing chip for rapid detection of the dengue virus

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Abstract

The enzyme-linked immunosorbent assay (ELISA) is widely used in medical diagnostics. In order to reduce the diagnosis time and to lower the consumption of sample/reagents in an ELISA assay, a suction-type, automatic, pneumatically-driven microfluidic chip has been designed and fabricated in this study. The microfluidic chip integrates a multi-functional micro-transport/mixing unit, for transporting metering and mixing of samples and reagents in order to automatically perform the entire ELISA protocol. A new surface modification has been adopted which allows for a high processing capacity. The detection sensitivity for the dengue virus is found to be 101 PFU/ml, which is much better than a conventional ELISA assay (103 PFU/ml). The entire assay time is only 30 min, which is much faster than with 96-well microtiter plates (4 h). The consumed sample and reagent volume is only 12 μl, which is less than a conventional assay (100 μl). The development of this microfluidic chip may be promising for other immunosensing applications.

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Abbreviations

BSA:

Bovine serum albumin

CCD:

Charge-coupled device

DI:

Deionized

DV:

Dengue virus

ELISA:

Enzyme-linked immunosorbent assay

EMV:

Electromagnetic valve

EV:

Enterovirus

GA:

Glutaraldehyde

HAU:

Hemagglutination unit

HBV:

Hepatitis B virus

HRP:

Horse reddish peroxidase

IgG:

Immunoglobulin G

IU:

International unit

LBL:

Layer-by-layer

OD:

Optical density

PAA:

Poly(acrylic acid)

PBS:

Phosphate buffer saline

PDMS:

Polydimethylsiloxane

PEI:

Poly(ethyleneimine)

PEM:

Polyelectrolyte multilayer

PFU:

Plaque forming unit

Pluronic P123:

Poly (ethylene oxide)-poly (propylene oxide)-poly (ethylene oxide) triblock copolymer

RT-PCR:

Reverse-transcription polymerase chain reaction

TMB:

Tetramethylbenzidin

UV:

Ultraviolet

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Acknowledgements

The authors gratefully acknowledge the financial support provided to this study by the National Science Council in Taiwan (NSC 98-2627-B-006-006; NSC 98-2120-M-006-001).

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Authors and Affiliations

  1. Department of Engineering Science, National Cheng Kung University, Tainan, 701, Taiwan

    Tze-Bin Huang

  2. Department of Chemistry, National Cheng Kung University, Tainan, 701, Taiwan

    Chih-Chia Huang & Chen-Sheng Yeh

  3. Department of Microbiology and Immunology, National Cheng Kung University, Tainan, 701, Taiwan

    Chen-Hsun Weng & Huan-Yao Lei

  4. Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, 30013, Taiwan

    Gwo-Bin Lee

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  1. Chen-Hsun Weng
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  2. Tze-Bin Huang
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  3. Chih-Chia Huang
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  4. Chen-Sheng Yeh
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  5. Huan-Yao Lei
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Correspondence to Gwo-Bin Lee.

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Weng, CH., Huang, TB., Huang, CC. et al. A suction-type microfluidic immunosensing chip for rapid detection of the dengue virus. Biomed Microdevices 13, 585–595 (2011). https://doi.org/10.1007/s10544-011-9529-3

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