Gorgannezhad Lena, Sreejith Kamalalayam Rajan, Christie Melody, Jin Jing, Ooi Chin Hong, Katouli Mohammad, Stratton Helen, Nguyen Nam-Trung
Queensland Micro- and Nanotechnology Centre, Nathan Campus, Griffith University, 170 Kessels Road, Brisbane, QLD 4111, Australia.
School of Environment and Science, Nathan Campus, Griffith University, 170 Kessels Road, Brisbane, QLD 4111, Australia.
Micromachines (Basel). 2020 Aug 7;11(8):761. doi: 10.3390/mi11080761.
Multiplex polymerase chain reaction (PCR) is an effective tool for simultaneous detection of target genes. Nevertheless, their use has been restricted due to the intrinsic interference between primer pairs. Performing several single PCRs in an array format instead of a multiplex PCR is a simple way to overcome this obstacle. However, there are still major technical challenges in designing a new generation of single PCR microreactors with a small sample volume, rapid thermal cycling, and no evaporation during amplification. We report a simple and robust core-shell bead array for a series of single amplifications. Four core-shell beads with a polymer coating and PCR mixture were synthesized using liquid marble formation and subsequent photo polymerization. Each bead can detect one target gene. We constructed a customised system for thermal cycling of these core-shell beads. Phylogrouping of the strains was carried out based on the fluorescent signal of the core-shell beads. This platform can be a promising alternative for multiplex nucleic acid analyses due to its simplicity and high throughput. The platform reported here also reduces the cycling time and avoids evaporation as well as contamination of the sample during the amplification process.
多重聚合酶链反应（PCR）是同时检测靶基因的有效工具。然而，由于引物对之间的内在干扰，其应用受到了限制。以阵列形式进行多个单重PCR而非多重PCR是克服这一障碍的简单方法。然而，设计新一代具有小样本量、快速热循环且扩增过程中无蒸发的单重PCR微反应器仍存在重大技术挑战。我们报道了一种用于一系列单重扩增的简单且稳健的核壳珠阵列。利用液滴形成及随后的光聚合反应合成了四种带有聚合物涂层和PCR混合物的核壳珠。每个珠子可检测一个靶基因。我们构建了一个用于这些核壳珠热循环的定制系统。基于核壳珠的荧光信号对菌株进行系统发育分组。该平台因其简单性和高通量，可能成为多重核酸分析的一个有前景的替代方案。本文报道的平台还缩短了循环时间，避免了扩增过程中样本的蒸发以及污染。
