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Mkit: A mobile nucleic acid assay based on a chitosan-modified minimalistic microfluidic chip (CM3-chip) and smartphone
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2023-03-15 , DOI: 10.1016/j.aca.2023.341030
Ke Yang 1 , Jingyu Pan 2 , Guoqing Deng 1 , Changyi Hua 1 , Cancan Zhu 1 , Yong Liu 1 , Ling Zhu 1
Affiliation  

Mobile sensing enabled by MS2 technology, which integrates microfluidic and smartphone components, has seen many applications in recent years. In this direction, we developed an MS2 platform (an integrated kit) for nucleic acid assay, which included a chitosan-modified minimalistic microfluidic chip (CM3-chip), a smartphone-based fluorescence detector (SF-detector), an APP for imaging and analysis, reagents, and accessories. Once the lysed sample was loaded into the CM3-chip modified by 1% concentration and 200–260 kDa molecular weight of chitosan, the following assay can be completed in approximately 1 h. The Mkit can detect 3 × 10° copies μL−1 of plasmid DNA and its polymerase chain reaction (PCR) efficiency was 96.8%. The CM3-chip equipped for the Mkit can enrich nucleic acid from the pH = 5 of lysis buffer, instead of using conventional adsorption mediums such as the magnetic beads and silica gel membranes, which could result in unexpected impurity residuals and tedious cleaning operations. In addition, the performance of the Mkit equipped with the pristine chip was demonstrated to perform poorer than that coupled with the CM3-chip in which the enriched nucleic acid can be all used for “in-situ PCR”. The universality, selectivity, and user-friendliness of the Mkit were also validated. We finally demonstrated the feasibility of the Mkit for testing artificially prepared infected samples. H5N6 and IAV-infected saliva samples provided the limits of detection of 5 × 102 copies mL−1 and 3.24 × 102 copies mL−1 per chamber, respectively. The streamlined assay and compact device should enable the great potential of the Mkit in research and potential diagnostic uses.



中文翻译:

Mkit:基于壳聚糖修饰的简约微流控芯片(CM3 芯片)和智能手机的移动核酸检测

集成了微流体和智能手机组件的 MS 2技术实现的移动传感近年来得到了许多应用。在这个方向上,我们开发了用于核酸检测的MS 2平台(集成套件),其中包括壳聚糖修饰的简约微流控芯片(CM 3芯片)、基于智能手机的荧光检测器(SF 检测器)、APP用于成像和分析、试剂和附件。一旦将裂解的样品加载到由 1% 浓度和 200–260 kDa 分子量的壳聚糖修饰的 CM 3芯片中,以下测定可在大约 1 小时内完成。Mkit 可以检测 3 × 10° 拷贝 μL −1质粒 DNA 及其聚合酶链反应 (PCR) 效率为 96.8%。Mkit配备的CM 3芯片可以从pH = 5的裂解缓冲液中富集核酸,而不是使用传统的吸附介质,如磁珠和硅胶膜,这可能会导致意想不到的杂质残留和繁琐的清洁操作。此外,配备原始芯片的 Mkit 的性能被证明比与 CM 3芯片耦合的性能更差,其中富集的核酸可以全部用于“原位”聚合酶链反应”。Mkit 的通用性、选择性和用户友好性也得到了验证。我们最终证明了 Mkit 用于测试人工制备的感染样本的可行性。H5N6 和 IAV 感染的唾液样本分别提供了每个腔室 5 × 10 2拷贝 mL -1和 3.24 × 10 2拷贝 mL -1的检测限。流线型化验和紧凑型设备应使 Mkit 在研究和潜在诊断用途方面具有巨大潜力。

更新日期:2023-03-20
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