Heater Integrated Lab-on-a-Chip Device for Rapid HLA Alleles Amplification towards Prevention of Drug Hypersensitivity.

Shah Mukim Uddin,Abkar Sayad,Jianxiong Chan,Duc Hau Huynh,Patrick Kwan,Efstratios Skafidas

doi:10.3390/s21103413

Abstract

HLA-B*15:02 screening before administering carbamazepine is recommended to prevent life-threatening hypersensitivity. However, the unavailability of a point-of-care device impedes this screening process. Our research group previously developed a two-step HLA-B*15:02 detection technique utilizing loop-mediated isothermal amplification (LAMP) on the tube, which requires two-stage device development to translate into a portable platform. Here, we report a heater-integrated lab-on-a-chip device for the LAMP amplification, which can rapidly detect HLA-B alleles colorimetrically. A gold-patterned micro-sized heater was integrated into a 3D-printed chip, allowing microfluidic pumping, valving, and incubation. The performance of the chip was tested with color dye. Then LAMP assay was conducted with human genomic DNA samples of known HLA-B genotypes in the LAMP-chip parallel with the tube assay. The LAMP-on-chip results showed a complete match with the LAMP-on-tube assay, demonstrating the detection system’s concurrence.

Highlights

The loop-mediated isothermal amplification (LAMP) amplicon hybridizes with the DNA probes immobilized on the interdigitated electrode (IDE)-based biosensor surface that act as the mono-allelic determinant of an human leukocyte antigen (HLA)-B*15:02 LAMP amplicon [19]
The LAMP-chip was tested with the color dye to assess the microfluidic operation, incubation
We have developed a LAMP-chip to amplify the HLA-B alleles, which is the first part of the two-step HLA-B*15:02 detection process

Summary

Introduction

Aromatic ring structured antiepileptic drugs (AEDs) such as phenytoin (PHT), lamotrigine (LTG), and carbamazepine (CBZ) are the most prevalent sources of severe cutaneous adverse drug reactions [1,2]. These range from benign to severe reactions, including drug reactions with eosinophilia, systemic symptoms (DRESS), acute generalized exanthematous pustulosis (AGEP), toxic epidermal necrolysis (TEN), and Stevens–Johnson syndrome (SJS) [3,4]. Pharmacogenetic studies have discovered genetic associations between antiepileptic drugs-induced cutaneous adverse drug reactions and the human leukocyte antigen (HLA) alleles. Pre-treatment HLA genotyping is recommended to prevent carbamazepine induced SJS/TEN. Several challenging factors need to be considered in the implementation of this recommendation [15]

Methods

Results

Discussion

Conclusion