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Biosensors
Special Issues
CRISPR-Cas Based Molecular Diagnostics and Their Applications
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CRISPR-Cas Based Molecular Diagnostics and Their Applications

A special issue of Biosensors (ISSN 2079-6374). This special issue belongs to the section "Nano- and Micro-Technologies in Biosensors".

Deadline for manuscript submissions: closed (31 December 2023) | Viewed by 11456

Special Issue Editors

Prof. Dr. Xiong Ding

E-Mail Website
Guest Editor
Department of Nutrition and Food Hygiene, School of Public Health, Southeast University, Nanjing 210009, China
Interests: molecular diagnostics; nucleic acid detection; biosensing
Special Issues, Collections and Topics in MDPI journals
Prof. Dr. Jin Wang

E-Mail Website
Guest Editor
Department of Nutrition and Food Hygiene, School of Public Health, Southeast University, Nanjing 210009, China
Interests: food allergy; protein structure analysis; novel food processing techniques; nutrition and human health
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The importance of the molecular diagnosis of diseases has been emphasized during the ongoing COVID-19 pandemic. Rapid and accurate molecular diagnostics are therefore highly desired to effectively prevent and control diseases, particularly emerging infectious diseases. Toward this end, a new generation of molecular diagnostics is formed by coupling the CRISPR-Cas systems. CRISPR-Cas-based diagnostics elevate the detection speed, sensitivity, and specificity, streamlining the molecular diagnostics towards point-of-care or on-site testing with high accuracy and reliability. In addition to underlying design of reaction systems, the combination with sensor chips and rapid sample processing expands the detection throughput and facilitates the development of miniaturized sensing devices with high integration. 

The aim of this Special Issue is to collect new forms and applications of CRISPR-Cas-based diagnostics with improvements in the analytical performance (speed, sensitivity, specificity, and selectivity), detection throughput (multiplexed), multifunctional integration (integrating sample preparation or in a sample-in–answer-out format), and miniaturization of sensing prototypes (microscale or portability). The new forms may involve novel assay principles, the employment of unique probes or dyes, the implementation of new CRISPR-Cas systems (new crRNA/sgRNA designs or new Cas nucleases), and the combination with plasmonic/nanoplasmonic biosensors, new-material biosensors, lab-on-a-chip biosensors, and others. Regarding real sample analysis, the studies should fully illustrate the advantages and disadvantages of the newly established CRISPR-Cas-based diagnostics in comparison with current standard methods. The applications include, but are not limited to, pathogen detection, clinical diagnostics, therapy evaluation, environmental monitoring, food safety/hygiene, and the health management of pasturage aquiculture. One-pot, one-step CRISPR-Cas-based diagnostics with simplified detection procedures, as well as digital quantitation applications using microchambers or droplets are highly anticipated. Apart from the biological molecules of nucleic acids and proteins, we are also collecting manuscripts describing the new CRISPR-Cas-based diagnosis of nonbiological molecules such as heavy metal ions, organic small molecules, metabolites, and toxicants.

Prof. Dr. Xiong Ding
Prof. Dr. Jin Wang
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Biosensors is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • CRISPR-Cas systems
  • CRISPR-based diagnostics
  • point-of-care biosensors
  • digital quantitation
  • lab-on-a-chip
  • microscale detection systems
  • multifunctional integration
  • microfluidic diagnostics
  • food adulteration detection
  • food safety detection

Published Papers (3 papers)

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Research

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12 pages, 1308 KiB  
Article
Comparison of Single-Stranded DNA Probes Conjugated with Magnetic Particles for Trans-Cleavage in Cas12a-Based Biosensors
by Aleksandr V. Ivanov, Irina V. Safenkova, Anatoly V. Zherdev, Yi Wan and Boris B. Dzantiev
Biosensors 2023, 13(7), 700; https://doi.org/10.3390/bios13070700 - 01 Jul 2023
Cited by 3 | Viewed by 3545
Abstract
Biosensors based on endonuclease Cas12 provide high specificity in pathogen detection. Sensitive detection using Cas12-based assays can be achieved using trans-cleaved DNA probes attached to simply separated carriers, such as magnetic particles (MPs). The aim of this work was to compare polyA, polyC, [...] Read more.
Biosensors based on endonuclease Cas12 provide high specificity in pathogen detection. Sensitive detection using Cas12-based assays can be achieved using trans-cleaved DNA probes attached to simply separated carriers, such as magnetic particles (MPs). The aim of this work was to compare polyA, polyC, and polyT single-stranded (ss) DNA with different lengths (from 10 to 145 nt) as trans-target probes were immobilized on streptavidin-covered MPs. Each ssDNA probe was labeled using fluorescein (5′) and biotin (3′). To compare the probes, we used guide RNAs that were programmed for the recognition of two bacterial pathogens: Dickeya solani (causing blackleg and soft rot) and Erwinia amylovora (causing fire blight). The Cas12 was activated by targeting double-stranded DNA fragments of D. solani or E. amylovora and cleaved the MP–ssDNA conjugates. The considered probes demonstrated basically different dependencies in terms of cleavage efficiency. PolyC was the most effective probe when compared to polyA or polyT probes of the same length. The minimal acceptable length for the cleavage follows the row: polyC < polyT < polyA. The efficiencies of polyC and polyT probes with optimal length were proven for the DNA targets’ detection of D. solani and E. amylovora. The regularities found can be used in Cas12a-based detection of viruses, bacteria, and other DNA/RNA-containing analytes. Full article
(This article belongs to the Special Issue CRISPR-Cas Based Molecular Diagnostics and Their Applications)
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11 pages, 2204 KiB  
Article
CRISPR/Cas12a-Assisted Dual Visualized Detection of SARS-CoV-2 on Frozen Shrimps
by Siwenjie Qian, Yanju Chen, Xiaofu Wang, Tingzhang Wang, Yang Che, Jian Wu, Zhangying Ye and Junfeng Xu
Biosensors 2023, 13(1), 138; https://doi.org/10.3390/bios13010138 - 14 Jan 2023
Cited by 2 | Viewed by 1760
Abstract
Given the possibility that food contaminated with SARS-CoV-2 might become an infection source, there is an urgent need for us to develop a rapid and accurate nucleic acid detection method for SARS-CoV-2 in food to ensure food safety. Here, we propose a sensitive, [...] Read more.
Given the possibility that food contaminated with SARS-CoV-2 might become an infection source, there is an urgent need for us to develop a rapid and accurate nucleic acid detection method for SARS-CoV-2 in food to ensure food safety. Here, we propose a sensitive, specific, and reliable molecular detection method for SARS-CoV-2. It has a mechanism to control amplicon contamination. Swabs from spiked frozen shrimps were used as detection samples, which were processed by heating at 95 °C for 30 s. These preprocessed samples served as the templates for subsequent amplification. A colorimetric LAMP reaction was carried out to amplify both the SARS-CoV-2 target and the MS2 phage simultaneously in one tube. MS2 phage was detected by colorimetric LAMP as the internal control, while SARS-CoV-2 was detected with a CRISPR/Cas12a system. The fluorescence results could be visually detected with an ultraviolet lamp. Meanwhile, uracil was incorporated during the LAMP reaction to provide an amplicon contamination proof mechanism. This test could detect as low as 20 copies of SARS-CoV-2 in one reaction. Additionally, the detection could be finished in 45 min. The test only needs a heating block and an ultraviolet lamp, which shows the potential for field detection. Full article
(This article belongs to the Special Issue CRISPR-Cas Based Molecular Diagnostics and Their Applications)
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Review

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16 pages, 2411 KiB  
Review
Isothermal Amplification Technology for Disease Diagnosis
by Poramin Boonbanjong, Kiatnida Treerattrakoon, Wassa Waiwinya, Piyawat Pitikultham and Deanpen Japrung
Biosensors 2022, 12(9), 677; https://doi.org/10.3390/bios12090677 - 24 Aug 2022
Cited by 18 | Viewed by 5472
Abstract
Isothermal amplification (IA) is a nucleic acid amplification technology (NAAT) that has contributed significantly to the healthcare system. The combination of NAAT with a suitable detection platform resulted in higher sensitivity, specificity, and rapid disease diagnosis. Traditional NAAT, such as polymerase chain reaction [...] Read more.
Isothermal amplification (IA) is a nucleic acid amplification technology (NAAT) that has contributed significantly to the healthcare system. The combination of NAAT with a suitable detection platform resulted in higher sensitivity, specificity, and rapid disease diagnosis. Traditional NAAT, such as polymerase chain reaction (PCR), is widely applied in the general healthcare system but is rarely accessed in resource-limited hospitals. Some IA methods provide a rapid, sensitive, specific, and simple method for disease diagnosis. However, not all IA techniques have been regularly used in clinical applications because different biomarkers and sample types affect either the enzyme in the IA system or sample preparation. This review focuses on the application of some IA techniques that have been applied in the medical field and have the potential for use at points of care. Full article
(This article belongs to the Special Issue CRISPR-Cas Based Molecular Diagnostics and Their Applications)
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