Emerging ultrafast nucleic acid amplification technologies for next-generation molecular diagnostics

2019 ◽  
Vol 141 ◽  
pp. 111448 ◽  
Author(s):  
Sang Hun Lee ◽  
Seung-min Park ◽  
Brian N. Kim ◽  
Oh Seok Kwon ◽  
Won-Yep Rho ◽  
...  
2019 ◽  
Vol 116 (33) ◽  
pp. 16240-16249 ◽  
Author(s):  
Wei Ouyang ◽  
Jongyoon Han

Rapid and reliable detection of ultralow-abundance nucleic acids and proteins in complex biological media may greatly advance clinical diagnostics and biotechnology development. Currently, nucleic acid tests rely on enzymatic processes for target amplification (e.g., PCR), which have many inherent issues restricting their implementation in diagnostics. On the other hand, there exist no protein amplification techniques, greatly limiting the development of protein-based diagnosis. We report a universal biomolecule enrichment technique termed hierarchical nanofluidic molecular enrichment system (HOLMES) for amplification-free molecular diagnostics using massively paralleled and hierarchically cascaded nanofluidic concentrators. HOLMES achieves billion-fold enrichment of both nucleic acids and proteins within 30 min, which not only overcomes many inherent issues of nucleic acid amplification but also provides unprecedented enrichment performance for protein analysis. HOLMES features the ability to selectively enrich target biomolecules and simultaneously deplete nontargets directly in complex crude samples, thereby enormously enhancing the signal-to-noise ratio of detection. We demonstrate the direct detection of attomolar nucleic acids in urine and serum within 35 min and HIV p24 protein in serum within 60 min. The performance of HOLMES is comparable to that of nucleic acid amplification tests and near million-fold improvement over standard enzyme-linked immunosorbent assay (ELISA) for protein detection, being much simpler and faster in both applications. We additionally measured human cardiac troponin I protein in 9 human plasma samples, and showed excellent agreement with ELISA and detection below the limit of ELISA. HOLMES is in an unparalleled position to unleash the potential of protein-based diagnosis.


2015 ◽  
Vol 8 (1) ◽  
Author(s):  
Jianbing Qin ◽  
Jennifer N. Sanmann ◽  
Jeff S. Kittrell ◽  
Pamela A. Althof ◽  
Erin E. Kaspar ◽  
...  

2016 ◽  
Vol 12 (5) ◽  
pp. 386-396 ◽  
Author(s):  
Michael G. Mauk ◽  
Jinzhao Song ◽  
Yubing Tong ◽  
Haim H. Bau ◽  
Changchun Liu

2011 ◽  
Vol 135 (1) ◽  
pp. 83-91 ◽  
Author(s):  
Juan P. Olano ◽  
David H. Walker

Abstract Context—Molecular diagnostics continues to evolve very rapidly, and its impact in the diagnosis of infectious diseases is undeniable. Molecular tools have played a pivotal role in discovering and characterizing several emerging infectious agents and have now become the gold standard for the diagnosis of infectious diseases caused by fastidious or uncultivable agents. Multiple challenges still remain for the widespread use of cost-effective, validated, and commercially available molecular tools. Automated instruments capable of sample processing and multiplex nucleic acid amplification and postamplification analysis have already been approved by the US Food and Drug Administration (FDA) for use in the clinical setting. Nanobiotechnology is beginning to impact laboratory diagnostics in the clinical setting. Objective—To address current nucleic acid techniques used in the clinical laboratory for diagnosis of infectious diseases. FDA-approved tests are listed, as well as molecular techniques (amplification and postamplification analysis). A comprehensive list of emerging pathogens during the last 4 decades is also presented. Biosurveillance systems are discussed in the context of molecular tools. The rapidly evolving field of nanobiotechnology is briefly addressed. Data Sources—Original publications, major reviews, and book chapters were used to present a comprehensive, yet short, review of molecular diagnostics in infectious diseases. Conclusions—We will continue to witness an exponential growth of molecular techniques used for the initial diagnosis of infectious diseases. Molecular tools will also continue to have an impact on disease prognosis and response to therapeutic interventions. Automation, multiplexing, and miniaturization will continue to be driving forces in the development of new instruments.


2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Guoliang Huang ◽  
Qin Huang ◽  
Li Ma ◽  
Xianbo Luo ◽  
Biao Pang ◽  
...  

PLoS ONE ◽  
2011 ◽  
Vol 6 (5) ◽  
pp. e19738 ◽  
Author(s):  
Paul LaBarre ◽  
Kenneth R. Hawkins ◽  
Jay Gerlach ◽  
Jared Wilmoth ◽  
Andrew Beddoe ◽  
...  

2013 ◽  
Vol 58 (10) ◽  
pp. 1162-1168 ◽  
Author(s):  
Ryo Kubota ◽  
Paul Labarre ◽  
Bernhard H. Weigl ◽  
Yong Li ◽  
Paul Haydock ◽  
...  

2021 ◽  
Vol 34 (Suppl 1) ◽  
pp. 49-51
Author(s):  
Carmen Martín-Higuera ◽  
Irene Muñoz-Gallego ◽  
María Dolores Folgueira ◽  

The diagnosis of SARS-CoV-2 is based on the use of nucleic acid amplification tests (NAAT), especially rRT-PCR. The latter also allows us to quickly identify variants of concern. However, its use in follow-up of patients and the correlation between Ct value and the viability of the virus is controversial.


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