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Research articles

ScienceAsia 45(2019): 260-267 |doi: 10.2306/scienceasia1513-1874.2019.45.260


Dot-blot sandwich immunoassay with silver signal enhancement for simple amyloid beta protein fragment 1-42 detection


Soe Htut Wina, Prapimpun Wongchitratb, Sumana Kladsomboonc, Pornpatr A. Dharmasarojad, Amara Apiluxa,*

 
ABSTRACT:     Amyloid beta 1-42 (Aβ1-42) is a key pathophysiological hallmark of Alzheimer's disease (AD). Changes in biofluids of A? have been associated with AD risk. Herein, a dot-blot sandwich immunoassay was developed for simple detection of the Aβ1-42 using gold nanoparticles as signal label. The results were easily observed by naked eye, and an image processing program provided semi-quantitation. Furthermore, addition of silver enhancer to the assay further improved sensitivity of the measurement. The proposed assay for Aβ1-42 had a linear range of 1-200 μg/ml, with a detection limit of 1 μg/ml and 0.3 μg/ml by naked eye and calculation, respectively. Performance of the developed assay was assessed in analyses of spiked plasma samples. The recoveries of Aβ1-42 were in the range of 112-158%, indicating that the developed assay is practical and reliable. The proposed method provides a low cost and simple assay for Aβ1-42 measurement.

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a Department of Clinical Chemistry, Faculty of Medical Technology, Mahidol University, Salaya, Nakhon Pathom 73170 Thailand
b Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Salaya, Nakhon Pathom 73170 Thailand
c Faculty of Medical Technology, Mahidol University, Salaya, Nakhon Pathom 73170 Thailand
d Stroke and Neurodegenerative Diseases Research Unit, Faculty of Medicine, Thammasat University, Klong Luang, Pathumthani 12120 Thailand

* Corresponding author, E-mail: amara.apl@mahidol.edu

Received 29 Dec 2018, Accepted 21 Jun 2019