Industry 4.0 Convergence Bionics Engineering, Pukyong National University, Busan 48513, Republic of Korea.
School of Electrical Engineering, Pukyong National University, Busan 48513, Republic of Korea.
Biosensors (Basel). 2024 Mar 20;14(3):152. doi: 10.3390/bios14030152.
Accurate sample heating is vital for nucleic acid extraction and amplification, requiring a sophisticated thermal cycling process in nucleic acid detection. Traditional molecular detection systems with heating capability are bulky, expensive, and primarily designed for lab settings. Consequently, their use is limited where lab systems are unavailable. This study introduces a technique for performing the heating process required in molecular diagnostics applicable for point-of-care testing (POCT), by presenting a method for crafting customized heaters using freely patterned nichrome (NiCr) wire. This technique, fabricating heaters by arranging protrusions on a carbon black-polydimethylsiloxane (PDMS) cast and patterning NiCr wire, utilizes cost-effective materials and is not constrained by shape, thereby enabling customized fabrication in both two-dimensional (2D) and three-dimensional (3D). To illustrate its versatility and practicality, a 2D heater with three temperature zones was developed for a portable device capable of automatic thermocycling for polymerase chain reaction (PCR) to detect () O157:H7 pathogen DNA. Furthermore, the detection of the same pathogen was demonstrated using a customized 3D heater surrounding a microtube for loop-mediated isothermal amplification (LAMP). Successful DNA amplification using the proposed heater suggests that the heating technique introduced in this study can be effectively applied to POCT.
准确的样本加热对于核酸提取和扩增至关重要,这需要在核酸检测中进行复杂的热循环过程。具有加热功能的传统分子检测系统体积庞大、价格昂贵,主要设计用于实验室环境。因此,在实验室系统不可用时,其使用受到限制。本研究通过提出一种使用自由图案化的镍铬(NiCr)线制作定制加热器的方法,为适用于即时检测(POCT)的分子诊断中的加热过程提供了一种技术。该技术通过在碳黑-聚二甲基硅氧烷(PDMS)铸型上布置凸起并对 NiCr 线进行图案化来制作加热器,利用了具有成本效益的材料,不受形状限制,从而能够在二维(2D)和三维(3D)中进行定制制作。为了说明其多功能性和实用性,开发了一种具有三个温度区的 2D 加热器,用于能够自动进行聚合酶链反应(PCR)热循环的便携式设备,以检测 () O157:H7 病原体 DNA。此外,还使用环绕微管的定制 3D 加热器展示了对同一病原体的检测,用于环介导等温扩增(LAMP)。使用所提出的加热器成功进行了 DNA 扩增,表明本研究中引入的加热技术可以有效地应用于 POCT。