Disposable silicon-based all-in-one micro-qPCR for rapid on-site detection of pathogens.

Disposable silicon-based all-in-one micro-qPCR for rapid on-site detection of pathogens.
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一次性硅的多合一微QPCR,用于快速现场检测病原体。

DOI:
10.1038/s41467-020-19911-6
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发表时间:
2020-12-02
影响因子:
16.6
通讯作者:
Güder F
Güder F
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Nunez-Bajo E;Silva Pinto Collins A;Kasimatis M;Cotur Y;Asfour T;Tanriverdi U;Grell M;Kaisti M;Senesi G;Stevenson K;Güder F

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在处理高传染性病原体时,快速筛查和低成本诊断在选择正确的干预过程中起着至关重要的作用。如果致病因子没有有效的治疗方法,如新型冠状病毒SARS-CoV-2,并且没有表现出或类似于其他常见感染的症状,这一点尤其重要。在这里,我们报告了一种基于一次性硅的集成式需要点传感器(TriSilix),用于实时定量检测病原体特有的核酸序列。TriSilix可以在标准实验室中以晶片规模生产(10 × 10 × 0.65 mm大小的37个芯片可以在7 h内生产出来,每个器件的成本约为0.35美元)。通过实时荧光定量聚合酶链式反应可以定量检测到禽类副结核分枝杆菌基因组DNA的563 片段,在第35个循环中的检测限为20 Fg,相当于一种细菌。利用TriSilix,我们还从SARS-CoV-2(1 PG)中检测到了对2003年SARS-CoV具有高度特异性的cDNA.设计针对核酸的高效、快速和低成本的诊断技术仍然是一个挑战。在这里,作者提出了一种在标准湿实验室中生产的一次性硅基集成需要点传感器,能够对病原体特定的核酸序列进行化学放大和实时定量检测。
Rapid screening and low-cost diagnosis play a crucial role in choosing the correct course of intervention when dealing with highly infectious pathogens. This is especially important if the disease-causing agent has no effective treatment, such as the novel coronavirus SARS-CoV-2, and shows no or similar symptoms to other common infections. Here, we report a disposable silicon-based integrated Point-of-Need transducer (TriSilix) for real-time quantitative detection of pathogen-specific sequences of nucleic acids. TriSilix can be produced at wafer-scale in a standard laboratory (37 chips of 10 × 10 × 0.65 mm in size can be produced in 7 h, costing ~0.35 USD per device). We are able to quantitatively detect a 563 bp fragment of genomic DNA of Mycobacterium avium subspecies paratuberculosis through real-time PCR with a limit-of-detection of 20 fg, equivalent to a single bacterium, at the 35th cycle. Using TriSilix, we also detect the cDNA from SARS-CoV-2 (1 pg) with high specificity against SARS-CoV (2003). Designing efficient, rapid and low-cost diagnostic technologies targeting nucleic acids remains a challenge. Here the authors present a disposable silicon-based integrated Point-of-Need transducer produced in a standard wet lab and able to chemically-amplify and detect pathogen-specific sequences of nucleic acids quantitatively in real-time.
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