Design and Implementation of Experimental Consumable Management Device for the Automatic Nucleic Acid Detection System

2016 ◽  
Vol 16 (7) ◽  
pp. 7069-7076
Author(s):  
Jun Yu ◽  
Zhu Chen ◽  
Chao Wang ◽  
Yana Hu ◽  
Hongming Dong ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Detection System ◽  
Nucleic Acid Detection ◽  
Design And Implementation

Design and Implementation of a High-Throughput Vibrating Module for Nucleic Acid Detection System

2020 ◽  
Vol 20 (4) ◽  
pp. 2165-2170
Author(s):  
Yanping Huang ◽  
Chao Wang ◽  
He Zhu ◽  
Zunliang Wang ◽  
Hui Chen ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
High Throughput ◽  
Detection System ◽  
Nucleic Acid Detection ◽  
Design And Implementation

scholarly journals Scanning single-molecule counting system for Eprobe with highly simple and effective approach

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243319
Author(s):  
Takeshi Hanami ◽  
Tetsuya Tanabe ◽  
Takuya Hanashi ◽  
Mitsushiro Yamaguchi ◽  
Hidetaka Nakata ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Single Molecule ◽  
Detection System ◽  
Signal To Noise Ratio ◽  
High Sensitivity ◽  
Nucleic Acid Detection ◽  
Digital Measurement ◽  
Excitation Light ◽  
Target Nucleic Acid ◽  
Fluorescent Molecules

Here, we report a rapid and ultra-sensitive detection technique for fluorescent molecules called scanning single molecular counting (SSMC). The method uses a fluorescence-based digital measurement system to count single molecules in a solution. In this technique, noise is reduced by conforming the signal shape to the intensity distribution of the excitation light via a circular scan of the confocal region. This simple technique allows the fluorescent molecules to freely diffuse into the solution through the confocal region and be counted one by one and does not require statistical analysis. Using this technique, 28 to 62 aM fluorescent dye was detected through measurement for 600 s. Furthermore, we achieved a good signal-to-noise ratio (S/N = 2326) under the condition of 100 pM target nucleic acid by only mixing a hybridization-sensitive fluorescent probe, called Eprobe, into the target oligonucleotide solution. Combination of SSMC and Eprobe provides a simple, rapid, amplification-free, and high-sensitive target nucleic acid detection system. This method is promising for future applications to detect particularly difficult to design primers for amplification as miRNAs and other short oligo nucleotide biomarkers by only hybridization with high sensitivity.

Mechanical Gripper Design and Force Analysis of Microplates for Automated High-Throughput Nucleic Acid Detection System

2020 ◽  
Vol 20 (3) ◽  
pp. 1401-1408 ◽  
Author(s):  
Chao Wang ◽  
Zhukang Guo ◽  
He Zhu ◽  
Nongyue He ◽  
Zhu Chen ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
High Throughput ◽  
Detection System ◽  
Nucleic Acid Detection ◽  
Force Analysis ◽  
Control Board ◽  
Absolute Position ◽  
Element Analysis ◽  
Position Data ◽  
Master Control

In the automated high-throughput nucleic acid detection system, we need to grip and transfer consumables frequently when carrying out multichannel nucleic acid detection. In order to ensure the efficiency of experiments and solve problems of the deflection and drop when transferring microplates, we design a self-locking mechanical gripper which consists of a rotary positioning module and a gripping module. The absolute position encoder fixed on the top of the stepper motor can collect the position data of the mechanical gripper in real time and send them to the master control board based on STM32 for processing, which ensures the accuracy of the movement of the mechanical gripper. We used SolidWorks to build models of the mechanical gripper and different microplates, and we carried on finite element analysis of microplates to find the suitable gripping position. Through the force analysis, we obtained the pressure distribution and the deformation of different microplates, and defined the effective gripping areas, which is important to the grip and transfer of microplates.

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 Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA Platform

2021 ◽  
Vol 12 ◽  
Author(s):  
Yang You ◽  
Pingping Zhang ◽  
Gengshan Wu ◽  
Yafang Tan ◽  
Yong Zhao ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Yersinia Pestis ◽  
Rapid Detection ◽  
Detection Method ◽  
Detection System ◽  
Point Of Care ◽  
High Sensitivity ◽  
Nucleic Acid Detection ◽  
Great Promise ◽  
High Background

The recent discovery of collateral cleavage activity of class-II clustered regularly interspaced short palindromic repeats–CRISPR-associated protein (CRISPR-Cas) makes CRISPR-based diagnosis a potential high-accuracy nucleic acid detection method. Colloidal gold-based lateral flow immunochromatographic assay (LFA), which has been combined with CRISPR/Cas-based nucleic detection, usually associates with drawbacks of relative high background and the subjectivity in naked-eye read-out of the results. Here, we developed a novel system composed of Cas12a-based nucleic acid detection and up-converting phosphor technology (UPT)-based LFA (UPT–LFA), termed Cas12a-UPTLFA. We further demonstrated the utility of this platform in highly sensitive and specific detection of Yersinia pestis, the causative agent of the deadly plague. Due to high infectivity and mortality, as well as the potential to be misused as bioterrorism agent, a culture-free, ultrasensitive, specific, and rapid detection method for Y. pestis has long been desired. By incorporating isothermal recombinase polymerase amplification, the Cas12a-UPTLFA we established can successfully detect genomic DNA of Y. pestis as low as 3 attomolar (aM) and exhibited high sensitivity (93.75%) and specificity (90.63%) for detection of spiked blood samples with a detection limit of 102 colony-forming unit per 100 μl of mouse blood. With a portable biosensor, Cas12a-UPTLFA assay can be operated easily by non-professional personnel. Taken together, we have developed a novel Cas12a-UPTLFA platform for rapid detection of Y. pestis with high sensitivity and specificity, which is portable, not expensive, and easy to operate as a point-of-care method. This detection system can easily be extended to detect other pathogens and holds great promise for on-site detection of emerging infectious pathogens.

scholarly journals A Novel Amplification-free SARS-CoV-2 Point-of-care Nucleic Acid Detection System based on Hybrid Capture Fluorescence Immunoassay

2020 ◽  
Author(s):  
Daming Wang ◽  
Shaogui He ◽  
Xiaohui Wang ◽  
Min Chen ◽  
Li Li ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Detection System ◽  
Point Of Care ◽  
Quantitative Polymerase Chain Reaction ◽  
Nucleic Acid Detection ◽  
Kappa Statistics ◽  
Fluorescence Immunoassay ◽  
Time Saving ◽  
Hybrid Capture ◽  
Test Kit

Abstract Herein we presented a novel, rapid and amplification-free SARS-CoV-2 nucleic acid detection system based on hybrid capture fluorescence immunoassay (HC-FIA) technology. The usage of the monoclonal antibody S9.6 in recognizing DNA-RNA double-stranded hybrids enabled the conversion of nucleic acid testing into immunofluorescence carrying on a simple lateral flow dipstick. The established HC-FIA also exhibited satisfactory sensitivity, specificity and great robustness. The clinical evaluation of HC-FIA kit and fluorescence reading device are further processed in three hospitals independently. The results of 734 samples from 670 subjects indicated high consistency between our HC-FIA and quantitative polymerase chain reaction based commercially available kit or clinical diagnosis according to Kappa statistics. Altogether, HC-FIA related method and commercial test kit show unparalleled advantages as time saving, amplification-free, high throughput and portable POCT molecular diagnosis, which facilitates its application as on-site Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid detection in epidemic prevention and control worldwide, especially during the outbreak.

Labelfree fully electronic nucleic acid detection system based on a field-effect transistor device

2004 ◽  
Vol 19 (12) ◽  
pp. 1723-1731 ◽  
Author(s):  
F. Uslu ◽  
S. Ingebrandt ◽  
D. Mayer ◽  
S. Böcker-Meffert ◽  
M. Odenthal ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Field Effect ◽  
Field Effect Transistor ◽  
Detection System ◽  
Nucleic Acid Detection ◽  
Effect Transistor

Improvement and Application of qPCR (Real-Time Quantitative Polymerase Chain Reaction) Data Processing Method for Home-Made Integrated Nucleic Acid Detection System

2020 ◽  
Vol 20 (12) ◽  
pp. 7369-7375
Author(s):  
Yile Fang ◽  
Pei Liao ◽  
Zhu Chen ◽  
Hui Chen ◽  
Yanqi Wu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Data Processing ◽  
Real Time ◽  
Detection System ◽  
Disease Diagnosis ◽  
Processing Algorithm ◽  
Clinical Samples ◽  
Nucleic Acid Detection ◽  
Qpcr Data ◽  
Data Processing Algorithm

Because it has many advantages such as rapidity and accuracy, nucleic acid detection is applied to infectious disease diagnosis more and more. An automatic integrated nucleic acid detection system based on real-time PCR is developed by our research group to conduct point-of-care testing of infectious pathogens. The home-made detection system collects fluorescence data in each PCR cycle through an integrated dual-channel fluorescence detection module and then real-time fluorescence curves are drawn by the software, which can tell the results of the diagnostics after some processing and analysis. However, owing to the disturbance of the environment or the imperfect of nucleic acid extraction before PCR, the fluorescence curves sometimes may contain several abnormal points. For the purpose of enhancing its ability to deal with these iffy curves and improve the accuracy of the testing results, in this study, the SDM-based qPCR data processing algorithm was studied and 11 groups of qPCR data that have different flaws from the clinical samples detected by this system were chosen to prove the practicability of the method. In comparison with the conventional threshold-based method, the Cq values calculated by the SDM-based method were more close to the actual values, meaning it can overcome the shortcomings of the conventional methods such as being unable to accommodate noise and being unable to avoiding abnormal data. With the improvement of this data processing algorithm, the stability of our system and the reliability and accuracy of the results are greatly improved.

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