scholarly journals CRISPR-Assisted DNA Detection, a novel dCas9-based DNA detection technique

2020 ◽  
Author(s):  
Xinhui Xu ◽  
Tao Luo ◽  
Jinliang Gao ◽  
Na Lin ◽  
Weiwei Li ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Room Temperature ◽  
Dna Detection ◽  
Point Of Care ◽  
Hpv Infection ◽  
Solid Support ◽  
Human Papillomaviruses ◽  
Nucleic Acid Detection ◽  
Detection Techniques ◽  
Signal Readout

AbstractNucleic acid detection techniques are always critical to diagnosis, especially in the background of the present COVID-19 pandemic. The simple and rapid detection techniques with high sensitivity and specificity are always urgently needed. However, the current nucleic acid detection techniques are still limited the traditional amplification and hybridization. To overcome the limitation, we here develop a CRISPR/Cas9-assisted DNA detection (CADD). In this detection, DNA sample is incubated with a pair of capture sgRNAs (sgRNAa and sgRNAb) specific to a target DNA, dCas9, a signal readout-related probe, and an oligo-coated solid support beads or microplate at room temperature for 15 min. During this incubation, the dCas9-sgRNA-DNA complex is formed and captured on solid support by the capture sequence of sgRNAa and the signal readout-related probe is captured by the capture sequence of sgRNAb. Finally the detection result is reported by a fluorescent or colorimetric signal readout. This detection was verified by detecting DNA of bacteria, cancer cell and virus. Especially, by designing a set of sgRNAs specific to 15 high-risk human papillomaviruses (HPVs), the HPV infection in 64 clinical cervical samples were successfully detected by the method. All detections can be finished in 30 minutes at room temperature. This detection holds promise for rapid on-the-spot detection or point-of-care testing (POCT).

Achieving room temperature DNA amplification by dialling in destabilization

2015 ◽  
Vol 51 (44) ◽  
pp. 9101-9104 ◽  
Author(s):  
B. Safeenaz Alladin-Mustan ◽  
Catherine J. Mitran ◽  
Julianne M. Gibbs-Davis
Keyword(s):  
Nucleic Acid ◽  
Room Temperature ◽  
Point Of Care ◽  
Dna Amplification ◽  
Heating Element ◽  
Nucleic Acid Sequences

The ability to amplify nucleic acid sequences at room temperature without the need for any heating element has been achieved, which has promise in bio-diagnostics employed at the point of care.

A novel room temperature nucleic acid detection method based on immobilization of adenosine-based molecular beacon

2014 ◽  
Vol 198 ◽  
pp. 194-200 ◽  
Author(s):  
Ting-E. Du ◽  
Xun Mao ◽  
Manyu Jin ◽  
Tian Zhang ◽  
Yi Zhang
Keyword(s):  
Nucleic Acid ◽  
Room Temperature ◽  
Molecular Beacon ◽  
Detection Method ◽  
Nucleic Acid Detection

Cobalt disulfide nanowires as an effective fluorescent sensing platform for DNA detection

2016 ◽  
Vol 4 (17) ◽  
pp. 2860-2863 ◽  
Author(s):  
Zhicai Xing ◽  
Lei Wang ◽  
Xiurong Yang
Keyword(s):  
Nucleic Acid ◽  
Hydrothermal Method ◽  
Dna Detection ◽  
Nucleic Acid Detection ◽  
Fluorescent Sensing ◽  
Sensing Platform ◽  
Cobalt Disulfide ◽  
First Time

Cobalt disulfide nanowires are synthesized in solution using a facile two-step hydrothermal method for the first time and applied as an effective sensing platform for nucleic acid detection.

scholarly journals Quality Management for Point-Of-Care Testing of Pathogen Nucleic Acids: Chinese Expert Consensus

2021 ◽  
Vol 11 ◽  
Author(s):  
Xi Mo ◽  
Xueliang Wang ◽  
Zhaoqin Zhu ◽  
Yuetian Yu ◽  
Dong Chang ◽  
...  
Keyword(s):  
Public Health ◽  
Nucleic Acid ◽  
Quality Management ◽  
Clinical Laboratory ◽  
Point Of Care ◽  
Conventional Polymerase Chain Reaction ◽  
Laboratory Medicine ◽  
Expert Consensus ◽  
Nucleic Acid Detection ◽  
Point Of Care Testing

COVID-19 continues to circulate globally in 2021, while under the precise policy implementation of China’s public health system, the epidemic was quickly controlled, and society and the economy have recovered. During the pandemic response, nucleic acid detection of SARS-CoV-2 has played an indispensable role in the first line of defence. In the cases of emergency operations or patients presenting at fever clinics, nucleic acid detection is required to be performed and reported quickly. Therefore, nucleic acid point-of-care testing (POCT) technology for SARS-CoV-2 identification has emerged, and has been widely carried out at all levels of medical institutions. SARS-CoV-2 POCT has served as a complementary test to conventional polymerase chain reaction (PCR) batch tests, thus forming an experimental diagnosis platform that not only guarantees medical safety but also improves quality services. However, in view of the complexity of molecular diagnosis and the biosafety requirements involved, pathogen nucleic acid POCT is different from traditional blood-based physical and chemical index detection. No guidelines currently exist for POCT quality management, and there have been inconsistencies documented in practical operation. Therefore, Shanghai Society of Molecular Diagnostics, Shanghai Society of Laboratory Medicine, Clinical Microbiology Division of Shanghai Society of Microbiology and Shanghai Center for Clinical Laboratory have cooperated with experts in laboratory medicine to generate the present expert consensus. Based on the current spectrum of major infectious diseases in China, the whole-process operation management of pathogen POCT, including its application scenarios, biosafety management, personnel qualification, performance verification, quality control, and result reporting, are described here. This expert consensus will aid in promoting the rational application and robust development of this technology in public health defence and hospital infection management.

scholarly journals A Chemical-Enhanced System for CRISPR-Based Nucleic Acid Detection

2021 ◽  
Author(s):  
Zihan Li ◽  
Wenchang Zhao ◽  
Shixin Ma ◽  
Zexu Li ◽  
Yingjia Yao ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Detection System ◽  
Point Of Care ◽  
Detection Sensitivity ◽  
Nucleic Acid Detection ◽  
Chemical Additives ◽  
Lateral Flow Strip ◽  
Viral Pathogens ◽  
Detection Systems ◽  
System Robustness

The CRISPR-based nucleic acid detection systems such as SHERLOCK, DETECTR and HOLMES have shown great potential for point-of-care testing of viral pathogens, especially in the context of COVID-19 pandemic. Here we optimize several key parameters of reaction chemistry and develop a Chemical Enhanced CRISPR Detection system for nucleic acid (termed CECRID). For the Cas12a/Cas13a-based signal detection phase, we determine buffer conditions and substrate range for optimal detection performance. By comparing several chemical additives, we find that addition of L-proline can secure or enhance Cas12a/Cas13a detection capability. For isothermal amplification phase with typical LAMP and RPA methods, inclusion of L-proline can also enhance specific target amplification as determined by CRISPR detection. Using SARS-CoV-2 pseudovirus, we demonstrate CECRID has enhanced detection sensitivity over chemical additive-null method with either fluorescence or lateral flow strip readout. Thus, CECRID provides an improved detection power and system robustness towards practical application of CRISPR-based diagnostics.

scholarly journals A Fast, Visual, and Instrument-free Platform Involving Rapid DNA Extraction, Chemical Heating, and Recombinase Aided Amplification for On-Site Nucleic Acid Detection

2021 ◽  
Vol 9 ◽  
Author(s):  
Xiao Fu Wang ◽  
Wen Qiang Chen ◽  
Jian Li Guo ◽  
Cheng Peng ◽  
Xiao Yun Chen ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Dna Extraction ◽  
Molecular Detection ◽  
Future Research ◽  
Nucleic Acid Detection ◽  
Valuable Insight ◽  
Detection Techniques ◽  
Detection Process ◽  
Novel Approach ◽  
Chemical Heating

The nucleic acid-based technique has been widely utilized in many fields including for on-site detection. However, traditional molecular detection techniques encounter limitations like relying on instruments, time consuming or complex operation, and cannot meet the demands of on-site testing. In this study, a rapid DNA extraction method (RDEM), recombinase aided amplification (RAA), and chemical heating packet (CHP) are integrated and termed as RRC platform for on-site detection of nucleic acid. For demonstration purposes, SHZD32-1 (a new transgenic soybean line from China) was detected using the novel platform to demonstrate its feasibility and capability for on-site detection. Using the RDEM, high-quality DNA appropriate for molecular detection was quickly extracted in 3–5 min. The heat energy generated by CHP was met the temperature requirements of RAA. Using the RRC platform, the whole detection process can be accomplished within only 30 min, and the results can be visually detected with glasses under blue light. No special or expensive instrument was needed for the detection process. This study provides a novel approach for on-site detection of nucleic acids besides providing valuable insight on related future research.

Magnetic colloids: preparation and biomedical applications

e-Polymers ◽  
2005 ◽  
Vol 5 (1) ◽  
Author(s):  
Abdelhamid Elaissari
Keyword(s):  
Nucleic Acid ◽  
Biomedical Applications ◽  
Colloidal Particles ◽  
Solid Support ◽  
Diagnostic Methods ◽  
Nucleic Acid Detection ◽  
Specific Detection ◽  
Biomedical Diagnosis ◽  
Biomedical Diagnostics ◽  
Limited Sensitivity

AbstractIncreasing interest has been focused on the preparation of magnetic latex particles and their use in biomedical diagnostics. All elaborated magnetic colloidal particles or latexes are principally used as a solid support of biomolecules involved in the specific capture of targeted biomolecules such as antigens in immunoassays (i.e., ELISA) and nucleic acid detection in ELOSA. Nowadays, the main objective in biomedical diagnosis is to enhance the sensitivity and to accelerate the procedure by elaborating new magnetic colloidal particles and diagnostic methods. The general problem in this field is related to the limited sensitivity. One possibility to partially solve it is to increase the concentration of captured targets (antigens, nucleic acids, etc.) before the specific detection step. Thus, the use of appropriate magnetic colloidal particles and methods for extraction and concentration of biomolecules is of extreme interest.

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