Peanut Detection Using Droplet Microfluidic Polymerase Chain Reaction Device

Shou-Yu Ma,An-Chong Chao,Chin-Chi Hsu,Yung-Sheng Lin,Yu-Cheng Chiang,Chia-Hsien Hsu,Jyh-Jian Chen

doi:10.1155/2019/4712084

Abstract

In this study, we integrated genetic detection for polymerase chain reaction (PCR) with microfluidics technology for the detection of peanut DNA. A cross-junction microchannel was used to induce emulsion droplets of water in oil for PCR on a chip. Compared with the single-phase flow, the emulsion droplet flow exhibited a 7.24% lower evaporation amount and prevented air bubble generation. PCR results of the droplet microfluidic PCR chip for peanut DNA fragment detection was verified by comparison with a commercial PCR thermal cycler and increased fluorescence intensity in SYBR Green reagent-based PCR. Moreover, PCR on the microfluidic PCR chip was successful for sesame, Salmonella spp., and Staphylococcus aureus. The droplet microfluidic PCR device developed in this study can be applied for peanut detection in the context of food allergy.

Highlights

Food allergy is a critical public health problem affecting children and adults [1]
Microfluidic polymerase chain reaction (PCR) devices can be classified into two types, namely, single-phase microfluidic PCR and droplet microfluidic PCR
This study developed a droplet microfluidic PCR device to amplify specific peanut DNA fragments for detection of foodborne allergens

Summary

Introduction

Allergy to peanuts is one of the most common food allergies [1,2,3]. Compared with macroscopic equivalents in polymerase chain reaction (PCR) systems, the microfluidic PCR device has several advantages, such as reduced sample and reagent consumption; these advantages enable inexpensive system operation and facilitate small thermal mass, low thermal inertia, and rapid heat transfer, improving the efficiency of PCR amplification [7]. Microfluidic PCR devices can be classified into two types, namely, single-phase microfluidic PCR and droplet microfluidic PCR. Droplet microfluidic systems using two immiscible fluids have emerged as a promising tool. These systems reduce analysis times, improve sensitivity, lower detection limits, increase high-throughput screening, and enhance operational flexibility [11,12,13]. Several studies of on-chip integration for droplet microfluidic PCR have been well reviewed [14]

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Conclusion