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Mkit:一种基于壳聚糖修饰的简约微流控芯片(CM芯片)和智能手机的移动核酸检测方法。

Mkit: A mobile nucleic acid assay based on a chitosan-modified minimalistic microfluidic chip (CM-chip) and smartphone.

作者信息

Yang Ke, Pan Jingyu, Deng Guoqing, Hua Changyi, Zhu Cancan, Liu Yong, Zhu Ling

机构信息

Anhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui, China.

Anhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui, China; Hefei Zhongke Yikangda Biomedical Co., LTD, Hefei, Anhui, China.

出版信息

Anal Chim Acta. 2023 May 1;1253:341030. doi: 10.1016/j.aca.2023.341030. Epub 2023 Mar 15.

Abstract

Mobile sensing enabled by MS technology, which integrates microfluidic and smartphone components, has seen many applications in recent years. In this direction, we developed an MS platform (an integrated kit) for nucleic acid assay, which included a chitosan-modified minimalistic microfluidic chip (CM-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 CM-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 of plasmid DNA and its polymerase chain reaction (PCR) efficiency was 96.8%. The CM-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 CM-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 × 10 copies mL and 3.24 × 10 copies mL per chamber, respectively. The streamlined assay and compact device should enable the great potential of the Mkit in research and potential diagnostic uses.

摘要

由微流控技术与智能手机组件集成而成的移动传感技术近年来已有诸多应用。在此方向上,我们开发了一种用于核酸检测的微流控平台(集成套件),其包括一个壳聚糖修饰的简约微流控芯片(CM芯片)、一个基于智能手机的荧光检测器(SF检测器)、一个用于成像和分析的应用程序、试剂及配件。一旦将裂解后的样品加载到经1%浓度、分子量为200 - 260 kDa的壳聚糖修饰的CM芯片中,后续检测大约可在1小时内完成。该M套件可检测3×10⁰拷贝/μL的质粒DNA,其聚合酶链反应(PCR)效率为96.8%。配备于M套件的CM芯片能够从pH = 5的裂解缓冲液中富集核酸,而非使用磁珠和硅胶膜等传统吸附介质,因为这些传统介质可能会导致意外的杂质残留和繁琐的清洗操作。此外,已证明配备原始芯片的M套件的性能不如配备CM芯片的M套件,在CM芯片中富集的核酸可全部用于“原位PCR”。该M套件的通用性、选择性和用户友好性也得到了验证。我们最终证明了该M套件用于检测人工制备的感染样本的可行性。H5N6和IAV感染的唾液样本每腔室的检测限分别为5×10拷贝/mL和3.24×10拷贝/mL。这种简化的检测方法和紧凑的设备应能使M套件在研究和潜在诊断用途方面具有巨大潜力。

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