Immobilization of DNA Aptamer on Quartz Crystal Microbalance

(QCM) Disc for Biosensing of Dengue Virus

Regina Aileen May V. Vergara, PhD1,2, Ma. Wennie Pauline C. Caelian1
1Gregor Mendel Research Laboratories, 2Department of Chemistry and Physics

University of San Agustin, General Luna Street, Iloilo City, Iloilo 5000, Philippines

INTRODUCTION
Distribution of Dengue, Worldwide, 2016 (WHO) ILOILO DENGUE CASES 2008-2017
14000

12000

Number of Cases
10000

131, 827 cases were 8000

recorded for the year 6000
2017
4000

2000

0
2008 2009 2010 2011 2012 2013 2014 2015 2016 2017
GOAL: Year
Characterize an immobilization of DNA aptamer on a Quartz Crystal Microbalance
(QCM) disc to develop a diagnostic biosensing device for early detection of dengue
virus.
METHODOLOGY RESULTS AND DISCUSSION
Preparation of Quartz Crystal Microbalance Discs

Blow-dried in Direct Attachment

Piranha solution Triple distilled Alcohol
nitrogen gas Method
water

Thiolated aptamer
immobilization
Direct Attachment Method Attachment using Streptavidin Method

Attachment using
Streptavidin Method

Avidin Immobilization Biotinylated aptamer

Thiolated Washing with triple distilled water immobilization
Incubation
aptamer solution
buffer

Scanning Electron Microscopy Results

8μm 3μm
(A) Gold surface with Thiolated aptamer, x 10000 (B) Gold surface with Biotynilated aptamer, x 25000
Minimal agitation at 80 rpm for 6 SEM results shows deposition of(A) Thiolated aptamer and (B) Biotynilated aptamer around
hours. the gold surface of the Quartz Crystal Microbalance (QCM) disc viewed at x10,000 and x
2500 magnification respectively.
Streptavidin solution
Washing with triple Thiolated DNA Aptamer
distilled water

Phosphate buffer, Biotynilated

pH 7.5 solution aptamer solution
(C) Thiolated DNA aptamer, x 100 (D) Thiolated DNA aptamer, x2700 (D) Thiolated DNA aptamer, x 8500

Thiolated DNA- aptamer immobilized on gold surface of Quartz Crystal Microbalance

Disc viewed in x 100, x2700 and x8,500 magnification.
Biotinylated DNA Aptamer

Washing with triple distilled water

(F) Biotnylated DNA aptamer, x100 (G) Biotnylated DNA aptamer, x 2,300

Well- distributed deposition around the Surface morphology of QCM disc

gold surface of QCM crystal reveals attachment of Biotinylated
Scanning Electron immobilized with Biotinylated DNA- DNA-aptamer in x2,300 magnification.
Microscopy Testing aptamer was seen in x100
magnification.

The gold surface of the CONCLUSIONS

QCM disc was subjected to
SEM testing to identify and
DNA –aptamer was successfully seen attached to the gold surface of
characterize its surface QCM disc of which this DNA-aptamer was found to bind to envelope
morphology. protein specifically ED-3 protein of the Dengue virus. This could pave the
way for the development of diagnostic sensor for early detection of
dengue virus.

ACKNOWLEDGMENTS
We would like to thank the Gregor Mendel Research Laboratories of the University of San Agustin, Iloilo for funding 1. REFERENCES
Chen; L., H.; Hsiao, W. H.; Lee, H. C.; Wu, S. C.; Cheng, J. W., Selection and Characterization of DNA Aptamers Targeting All Four

i
Serotypes of Dengue Viruses. PLOS ONE 2015, 1-13.
this research and also to Mr. Norwell Bautista of Southeast Asian Fisheries Development Center (SEAFDEC), Mr. 2. Min, K.; Cho, M.; Han, S. Y.; Shim, Y. B.; Ku, J.; Ban, C., A Simple and Direct Electrochemical Detection of Interferon- γ using its RNA and
DNA Aptamers. Biosensors and Bioelectronics 2008, 23, 1819-1824.
Gerhrd Perez and Ms. Jomelyn of Katrin Field Incorporated for the Scanning Electron Microscopy testing results. 3. Kim, Y. S., Jung, H. S., Matsuura, T., Lee, H. Y., Kawai, T., Gu, M. B., Electrochemical Detection of 17 β-estradiol using DNA Aptamer
Immobilized Gold Electrode Chip. Biosensors and Bioelectronics 2007, 22, 2525-2531
4. World Health Organization, 2016
5. Department of Health, 2017