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2nd International Conference on Artificial Intelligence and Smart Energy, ICAIS 2022 ; : 1714-1719, 2022.
Article in English | Scopus | ID: covidwho-1806905

ABSTRACT

COVID-19 is an infectious disease that causes serious pandemic. There are many means to contradict the outspread of the microbe and one such way is to sterilize the hands by washing with soap for 20 seconds. Despite washing hands regularly, there is a tendency to touch the contaminated surface in the tap and door handles. In order to make the hands contamination-free before entering the premise, a solution that a person will be allowed to enter only after hand washing has been implemented. Having an automatic faucet and door which will be controlled using the micro controller Arduino Nano wherein various sensors like IR sensor and other devices like servo motor, relay module and LCD display will be interfaced with the micro controller has been proposed. An IR sensor will be placed at the door that senses whether the person has washed his/her hands or not and grant access for the entry. Another IR sensor will be placed at the faucet and it will sense the person's hand and control the water flow accordingly. The proposed system will ensure whether people have washed their hands before entering their premises and grant access accordingly. © 2022 IEEE.

2.
Biosens Bioelectron ; 202: 113994, 2022 Apr 15.
Article in English | MEDLINE | ID: covidwho-1633350

ABSTRACT

The pandemic due to the outbreak of 2019 coronavirus disease (COVID-19) caused by novel severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has raised significant public health concerns. Rapid, affordable, and accurate diagnostic testing not only paves the way for the effective treatment of diseases, but also plays a crucial role in preventing the spreading of infectious diseases. Herein, a one-pot CRISPR/Cas13a-based visual biosensor was proposed and developed for the rapid and low-cost nucleic acid detection. By combining Cas13a cleavage and Recombinase Polymerase Amplification (RPA) in a one-pot reaction in a disposable tube-in-tube vessel, amplicon contamination could be completely avoided. The RPA reaction is carried out in the inner tube containing two hydrophobic holes at the bottom. After the completion of amplification reaction, the reaction solution enters the outer tube containing pre-stored Cas13a reagent under the action of centrifugation or shaking. Inner and outer tubes are combined to form an independent reaction pot to complete the nucleic acid detection without opening the lid. This newly developed nucleic acid detection method not only meets the need of rapid nucleic acid detection at home without the need for any specialized equipment, but also fulfils the requirement of rapid on-site nucleic acid detection with the aid of small automated instruments. In this study, CRISPR/Cas13a and CRISPR/Cas12a were used to verify the reliability of the developed one-pot nucleic acid detection method. The performance of the system was verified by detecting the DNA virus, i.e., African swine fever virus (ASFV) and the RNA virus, i.e., SARS-Cov-2. The results indicate that the proposed method possesses a limit of detection of 3 copy/µL. The negative and positive test results are consistent with the results of real-time fluorescence quantitative polymerase chain reaction (PCR), but the time required is shorter and the cost is lower. Thus, this study makes this method available in resource-limited areas for the purpose of large-scale screening and in case of epidemic outbreak.


Subject(s)
African Swine Fever Virus , Biosensing Techniques , COVID-19 , Nucleic Acids , Animals , CRISPR-Cas Systems , Humans , Nucleic Acid Amplification Techniques/methods , RNA, Viral/genetics , Reproducibility of Results , SARS-CoV-2 , Sensitivity and Specificity , Swine
3.
Biotechnol Bioeng ; 118(5): 2053-2066, 2021 05.
Article in English | MEDLINE | ID: covidwho-1092501

ABSTRACT

Polymerase chain reaction (PCR), a central technology for molecular diagnostics, is highly sensitive but susceptible to the risk of false positives caused by aerosol contamination, especially when an end-point detection mode is applied. Here, we proposed a solution by designing a clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 eraser strategy for eliminating potential contamination amplification. The CRISPR/Cas9 engineered eraser is firstly adopted into artpcr reverse-transcription PCR (RT-PCR) system to achieve contamination-free RNA detection. Subsequently, we extended this CRISPR/Cas9 eraser to the PCR system. We engineered conventional PCR primers to enable the amplified products to contain an implanted NGG (protospacer adjacent motif, PAM) site, which is used as a code for specific CRISPR/Cas9 recognition. Pre-incubation of Cas9/sgRNA with PCR mix leads to a selective cleavage of contamination amplicons, thus only the template DNA is amplified. The developed CRISPR/Cas9 eraser, adopted by both RT-PCR and PCR systems, showed high-fidelity detection of SARS-CoV-2 and African swine fever virus with a convenient strip test.


Subject(s)
CRISPR-Cas Systems , Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , African Swine Fever Virus/isolation & purification , Animals , Humans , RNA, Guide, Kinetoplastida , RNA, Viral/isolation & purification , SARS-CoV-2/isolation & purification , Swine
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