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Point-of-care DNA testing by automatically and sequentially performing extraction, amplification and identification in a closed-type cassette.
Sensors and Actuators B: Chemical ( IF 8.0 ) Pub Date : 2020-09-17 , DOI: 10.1016/j.snb.2020.128919
Tianhui Dong 1, 2 , Xueping Ma 2 , Nan Sheng 1, 2 , Xiemin Qi 2 , Yanan Chu 2 , Qinxin Song 1 , Bingjie Zou 2 , Guohua Zhou 1, 2, 3
Affiliation  

Nucleic acid detection is important for clinical diagnostics; however, it is challenging to perform genetic testing at the point-of-care due to the tedious steps involved in DNA extraction and the risk of cross-contamination from amplicons. To achieve a fully-automated and contamination-free nucleic acid detection, we propose a closed-type cassette system which enables the following steps to be operated automatically and sequentially: sample preparation based on magnetic beads, target amplification using multiplex polymerase chain reaction, and colorimetric detection of amplicons using a serial invasive reaction coupled with the aggregation of gold nanoparticle probes. The cassette was designed to be round and closed, and 10 targets in a sample could be simultaneously detected by the naked eye or using a spectrophotometer in the system. In addition, a cassette-driven device was fabricated to transfer reagents between wells, to control the temperature of each reaction, and to sense the colour in the detection wells. The cassette system was sensitive enough to detect 10 genotypes at 5 single nucleotide polymorphism sites related to the anticoagulant’s usage, by using a 0.5 μL blood sample. The accuracy of the system was evaluated by detecting 12 whole blood samples, and the results obtained were consistent with those obtained using pyrosequencing. The cassette is airtight and the whole system is fully automatic; the only manual operation is the addition of the sample to the cassette, performing point-of-care genetic testing in a sample-in/answer-out way.



中文翻译:

通过在封闭式暗盒中自动依次进行提取,扩增和鉴定来进行即时护理DNA测试。

核酸检测对临床诊断很重要;然而,由于涉及DNA提取的繁琐步骤以及扩增子交叉污染的风险,因此在现场进行基因检测具有挑战性。为了实现全自动且无污染的核酸检测,我们提出了一种封闭式盒式系统,该系统可自动进行以下步骤:基于磁珠的样品制备,使用多重聚合酶链反应的靶标扩增以及比色法检测扩增子,使用一系列侵袭性反应与金纳米粒子探针的聚集相结合。暗盒设计成圆形且封闭,可以用肉眼或系统中的分光光度计同时检测样品中的10个目标。另外,制造了盒式驱动装置以在孔之间转移试剂,控制每个反应的温度并感测检测孔中的颜色。该盒式系统足够灵敏,可以通过使用0.5μL血液样本在5个与抗凝剂使用相关的单核苷酸多态性位点检测10种基因型。该系统的准确性通过检测12个全血样品进行评估,所得结果与使用焦磷酸测序获得的结果一致。磁带盒是气密的,整个系统是全自动的。唯一的手动操作是将样品添加到盒中,以进样/出样的方式进行即时基因检测。并感应检测井中的颜色。该盒式系统足够灵敏,可以通过使用0.5μL血液样本在5个与抗凝剂使用相关的单核苷酸多态性位点检测10种基因型。该系统的准确性通过检测12个全血样品进行评估,所得结果与使用焦磷酸测序获得的结果一致。磁带盒是气密的,整个系统是全自动的。唯一的手动操作是将样品添加到盒中,以进样/出样的方式进行即时基因检测。并感应检测井中的颜色。该盒式系统足够灵敏,可以通过使用0.5μL血液样本在5个与抗凝剂使用相关的单核苷酸多态性位点检测10种基因型。该系统的准确性通过检测12个全血样品进行评估,所得结果与使用焦磷酸测序获得的结果一致。磁带盒是气密的,整个系统是全自动的。唯一的手动操作是将样品添加到盒中,以进样/出样的方式进行即时基因检测。该系统的准确性通过检测12个全血样品进行评估,所得结果与使用焦磷酸测序获得的结果一致。磁带盒是气密的,整个系统是全自动的。唯一的手动操作是将样品添加到盒中,以进样/出样的方式进行即时基因检测。该系统的准确性通过检测12个全血样品进行评估,所得结果与使用焦磷酸测序获得的结果一致。磁带盒是气密的,整个系统是全自动的。唯一的手动操作是将样品添加到盒中,以进样/出样的方式进行即时基因检测。

更新日期:2020-09-28
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