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Nucleic Acid Based Lateral Flow Strip Biosensor Via Competitive Binding For Possible Dengue Detection 2155 6210

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Nucleic Acid Based Lateral Flow Strip Biosensor Via Competitive Binding For Possible Dengue Detection 2155 6210
Yu et al., J Biosens Bioelectron 2012, 3:5 http://dx.doi.org/10.4172/2155-6210.1000128 Biosensors & Bioelectronics
Research Article

Open Access

Nucleic-Acid Based Lateral Flow Strip Biosensor via Competitive Binding for Possible Dengue Detection
Henson L. Lee Yu1, Christine Marie Montesa2, Nina Rosario L. Rojas1 and Erwin P. Enriquez1*
1
2

Department of Chemistry, Ateneo de Manila University, Quezon City, Philippines
JEOL Asia Ltd., Corporation Place, Singapore

Abstract
A low-cost, simple, rapid and selective nucleic-acid based lateral flow strip biosensor (LFSB) for possible dengue viral RNA detection is described in this study. The detection is based on competitive binding, where gold nanoparticles (AuNPs), with average size of ~10 nm confirmed using UV-Vis, TEM and AFM images, are used as visualizing agents. These are bioconjugated with DNA which competitively binds with its complementary strand either in the sample or in the test line of the LFSB. The detection scheme reduces the number of probes which effectively lowers the cost for the design of the test strip. The whole test took less than five minutes to complete and a red line signifies a negative result, while the absence of the line signifies a positive result. Quantification of the intensity of the red band reveals proportionality of the color to the amount of DNA present in the sample. The visual limit of detection of the LFSB is 10-7 M. It demonstrates selectivity in a blood matrix and selectivity over a synthetic
Influenza. This study brings us closer to an amplification-free, point-of-care method for dengue detection.

Keywords: Lateral flow strip biosensor; Dengue detection; Gold nanoparticles; Competitive binding assay

Introduction
A lateral flow strip biosensor (LFSB) is a type of sensor in which the elements are arranged in a strip of membrane and a test sample is allowed to elute through the membrane. The interpretation of the results will vary depending on the signal transduction


References: 1. Liu J, Cao Z, Lu Y (2009) Functional nucleic acid sensors. Chem Rev 109: 1948-1998. 2. Mao X, Ma Y, Zhang A, Zhang L, Zeng L, et al. (2009) Disposable nucleic acid biosensors based on gold nanoparticle probes and lateral flow strip 3. Glynou K, Ioannou PC, Christopoulos TK, Syriopoulou V (2003) Oligonucleotidefunctionalized gold nanoparticles as probes in a dry-reagent strip biosensor for DNA analysis by hybridization Sample 5. Baeumner AJ, Cohen RN, Miksic V, Min J (2003) RNA biosensor for the rapid 6. Yan Z, Zhou L, Zhao Y, Wang J, Huang L, et al. (2006) Rapid quantitative detection of Yersinia pestis by lateral-flow immunoassay and up-converting 7. Delmulle BS, De Saeger SM, Sibanda L, Barna-Vetro I, Van Peteghem CH (2005) Development of an immunoassay-based lateral flow dipstick for the relevant values (10-15-10-11M) for dengue viral DNA, a simple solidstate orthosilicate based [28] or magnetic nanoparticles-based (Kouassi 2006) pre-concentration step may be added, or stabilizers 8. Zhang G, Wang X, Zhi A, Bao Y, Yang Y, et al. (2008) Development of a lateral flow immunoassay strip for screening of sulfamonomethoxine residues 9. Huo Q (2007) A perspective on bioconjugated nanoparticles and quantum dots. 10. Posthuma-Trumpie GA, Korf J, van Amerongen A (2009) Lateral flow (immuno) assay: its strengths, weaknesses, opportunities and threats 11. Baeumner AJ, Schlesinger NA, Slutzki NS, Romano J, Lee EM, et al. (2002) Biosensor for dengue virus detection: sensitive, rapid, and serotype specific. 12. Poloni TR, Oliveira AS, Alfonso HL, Galvao LR, Amarilla AA, et al. (2010) Detection of dengue virus in saliva and urine by real time RT-PCR 14. Wu SJ, Lee EM, Putvatana R, Shurtliff RN, Porter KR, et al. (2001) Detection of dengue viral RNA using a nucleic acid sequence-based amplification assay. 15. Parida M, Horioke K, Ishida H, Dash PK, Saxena P, et al. (2005) Rapid detection and differentiation of dengue virus serotypes by a real-time reverse 16. Fry SR, Meyer M, Semple MG, Simmons CP, Sekaran SD, et al. (2011) The
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