Application of Molecular Diagnostic Technique of Nucleic Acid Analysis

A special issue of Biosensors (ISSN 2079-6374). This special issue belongs to the section "Biosensors and Healthcare".

Deadline for manuscript submissions: 23 July 2024 | Viewed by 12271

Special Issue Editor


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Guest Editor
Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, China
Interests: biosensors and biosystems; nucleic acid sensing chip; single-molecule identification; spatial omics sensing

Special Issue Information

Dear Colleagues,

Nucleic acids, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), are natural biopolymers composed of nucleotides that store, transmit, and express genetic information. Overexpressed or underexpressed as well as mutated nucleic acids have been implicated in many diseases. Therefore, nucleic acid diagnosis is extremely important.

Driven by pressing needs in clinical diagnosis and prevention of infectious diseases, nucleic acid diagnosis has evolved to be a rapidly advancing field. Currently, the reverse transcription polymerase chain reaction (RT-PCR) is the most widely used method. Other diagnostic molecular methods, including CRISPR-based assays, single-molecule identification, isothermal nucleic acid amplification methods, digital PCR, microarray assays, and next-generation sequencing (NGS), are promising alternatives.

The aim of this Special Issue is to invite authors to submit original research articles, short reports, or reviews that the progress to analyze nucleic acids, use nucleic acids as recognition probes, construct detection devices based on nucleic acids, and clinical applications of the different molecular diagnostic assays and other important fields.

Prof. Dr. Lianqun Zhou
Guest Editor

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Published Papers (2 papers)

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Research

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16 pages, 3377 KiB  
Article
A Critical Study on DNA Probes Attached to Microplate for CRISPR/Cas12 Trans-Cleavage Activity
by Konstantin M. Burkin, Aleksandr V. Ivanov, Anatoly V. Zherdev, Boris B. Dzantiev and Irina V. Safenkova
Biosensors 2023, 13(8), 824; https://doi.org/10.3390/bios13080824 - 17 Aug 2023
Cited by 1 | Viewed by 1244
Abstract
CRISPR/Cas12-based biosensors are emerging tools for diagnostics. However, their application of heterogeneous formats needs the efficient detection of Cas12 activity. We investigated DNA probes attached to the microplate surface and cleaved by Cas12a. Single-stranded (ss) DNA probes (19 variants) and combined probes with [...] Read more.
CRISPR/Cas12-based biosensors are emerging tools for diagnostics. However, their application of heterogeneous formats needs the efficient detection of Cas12 activity. We investigated DNA probes attached to the microplate surface and cleaved by Cas12a. Single-stranded (ss) DNA probes (19 variants) and combined probes with double-stranded (ds) and ssDNA parts (eight variants) were compared. The cleavage efficiency of dsDNA-probes demonstrated a bell-shaped dependence on their length, with a cleavage maximum of 50%. On the other hand, the cleavage efficiency of ssDNA probes increased monotonously, reaching 70%. The most effective ssDNA probes were integrated with fluorescein, antibodies, and peroxidase conjugates as reporters for fluorescent, lateral flow, and chemiluminescent detection. Long ssDNA probes (120–145 nt) proved the best for detecting Cas12a trans-activity for all of the tested variants. We proposed a test system for the detection of the nucleocapsid (N) gene of SARS-CoV-2 based on Cas12 and the ssDNA-probe attached to the microplate surface; its fluorescent limit of detection was 0.86 nM. Being united with pre-amplification using recombinase polymerase, the system reached a detection limit of 0.01 fM, thus confirming the effectiveness of the chosen ssDNA probe for Cas12-based biosensors. Full article
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Review

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23 pages, 4355 KiB  
Review
Principles and Applications of Loop-Mediated Isothermal Amplification to Point-of-Care Tests
by Jee-Woong Park
Biosensors 2022, 12(10), 857; https://doi.org/10.3390/bios12100857 - 10 Oct 2022
Cited by 24 | Viewed by 10235
Abstract
For the identification of nucleic acids, which are important biomarkers of pathogen-mediated diseases and viruses, the gold standard for NA-based diagnostic applications is polymerase chain reaction (PCR). However, the requirements of PCR limit its application as a rapid point-of-care diagnostic technique. To address [...] Read more.
For the identification of nucleic acids, which are important biomarkers of pathogen-mediated diseases and viruses, the gold standard for NA-based diagnostic applications is polymerase chain reaction (PCR). However, the requirements of PCR limit its application as a rapid point-of-care diagnostic technique. To address the challenges associated with regular PCR, many isothermal amplification methods have been developed to accurately detect NAs. Isothermal amplification methods enable NA amplification without changes in temperature with simple devices, as well as faster amplification times compared with regular PCR. Of the isothermal amplifications, loop-mediated isothermal amplification (LAMP) is the most studied because it amplifies NAs rapidly and specifically. This review describes the principles of LAMP, the methods used to monitor the process of LAMP, and examples of biosensors that detect the amplicons of LAMP. In addition, current trends in the application of LAMP to smartphones and self-diagnosis systems for point-of-care tests are also discussed. Full article
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