1.Development of a Recombinase-aided Amplification Combined With Lateral Flow Dipstick Assay for the Rapid Detection of the African Swine Fever Virus.
Jiang Shuai LI ; Yan Zhe HAO ; Mei Ling HOU ; Xuan ZHANG ; Xiao Guang ZHANG ; Yu Xi CAO ; Jin Ming LI ; Jing MA ; Zhi Xiang ZHOU
Biomedical and Environmental Sciences 2022;35(2):133-140
OBJECTIVE:
To establish a sensitive, simple and rapid detection method for African swine fever virus (ASFV) B646L gene.
METHODS:
A recombinase-aided amplification-lateral flow dipstick (RAA-LFD) assay was developed in this study. Recombinase-aided amplification (RAA) is used to amplify template DNA, and lateral flow dipstick (LFD) is used to interpret the results after the amplification is completed. The lower limits of detection and specificity of the RAA assay were verified using recombinant plasmid and pathogenic nucleic acid. In addition, 30 clinical samples were tested to evaluate the performance of the RAA assay.
RESULTS:
The RAA-LFD assay was completed within 15 min at 37 °C, including 10 min for nucleic acid amplification and 5 minutes for LFD reading results. The detection limit of this assay was found to be 200 copies per reaction. And there was no cross-reactivity with other swine viruses.
CONCLUSION
A highly sensitive, specific, and simple RAA-LFD method was developed for the rapid detection of the ASFV.
African Swine Fever/virology*
;
African Swine Fever Virus/isolation & purification*
;
Animals
;
Nucleic Acid Amplification Techniques/methods*
;
Recombinases/chemistry*
;
Sensitivity and Specificity
;
Swine
;
Viral Proteins/genetics*
2.A Longitudinal Survey for Genome-based Identification of SARS-CoV-2 in Sewage Water in Selected Lockdown Areas of Lahore City, Pakistan: A Potential Approach for Future Smart Lockdown Strategy.
Yaqub TAHIR ; Nawaz MUHAMMAD ; Z Shabbir MUHAMMAD ; A Ali MUHAMMAD ; Altaf IMRAN ; Raza SOHAIL ; A B Shabbir MUHAMMAD ; A Ashraf MUHAMMAD ; Z Aziz SYED ; Q Cheema SOHAIL ; B Shah MUHAMMAD ; Rafique SAIRA ; Hassan SOHAIL ; Sardar NAGEEN ; Mehmood ADNAN ; W Aziz MUHAMMAD ; Fazal SEHAR ; Hussain NADIR ; T Khan MUHAMMAD ; M Atique MUHAMMAD ; Asif ALI ; Anwar MUHAMMAD ; A Awan NABEEL ; U Younis MUHAMMAD ; A Bhattee MUHAMMAD ; Tahir ZARFISHAN ; Mukhtar NADIA ; Sarwar HUDA ; S Rana MAAZ ; Farooq OMAIR
Biomedical and Environmental Sciences 2021;34(9):729-733
3.Effects of Tanreqing Capsule on the negative conversion time of nucleic acid in patients with COVID-19: A retrospective cohort study.
Xing ZHANG ; Yan XUE ; Xuan CHEN ; Jia-Min WU ; Zi-Jian SU ; Meng SUN ; Lu-Jiong LIU ; Yi-Bao ZHANG ; Yi-le ZHANG ; Gui-Hua XU ; Miao-Yan SHI ; Xiu-Ming SONG ; Yun-Fei LU ; Xiao-Rong CHEN ; Wei ZHANG ; Qi CHEN
Journal of Integrative Medicine 2021;19(1):36-41
OBJECTIVE:
Traditional Chinese medicine plays a significant role in the treatment of the pandemic of coronavirus disease 2019 (COVID-19). Tanreqing Capsule (TRQC) was used in the treatment of COVID-19 patients in the Shanghai Public Health Clinical Center. This study aimed to investigate the clinical efficacy of TRQC in the treatment of COVID-19.
METHODS:
A retrospective cohort study was conducted on 82 patients who had laboratory-confirmed mild and moderate COVID-19; patients were treated with TRQC in one designated hospital. The treatment and control groups consisted of 25 and 57 cases, respectively. The treatment group was given TRQC orally three times a day, three pills each time, in addition to conventional Western medicine treatments which were also administered to the control group. The clinical efficacy indicators, such as the negative conversion time of pharyngeal swab nucleic acid, the negative conversion time of fecal nucleic acid, the duration of negative conversion of pharyngeal-fecal nucleic acid, and the improvement in the level of immune indicators such as T-cell subsets (CD3, CD4 and CD45) were monitored.
RESULTS:
COVID-19 patients in the treatment group, compared to the control group, had a shorter negative conversion time of fecal nucleic acid (4 vs. 9 days, P = 0.047) and a shorter interval of negative conversion of pharyngeal-fecal nucleic acid (0 vs. 2 days, P = 0.042). The level of CD3
CONCLUSION
Significant reductions in the negative conversion time of fecal nucleic acid and the duration of negative conversion of pharyngeal-fecal nucleic acid were identified in the treatment group as compared to the control group, illustrating the potential therapeutic benefits of using TRQC as a complement to conventional medicine in patients with mild and moderate COVID-19. The underlying mechanism may be related to the improved levels of the immune indicator CD3
Adult
;
Antiviral Agents/therapeutic use*
;
COVID-19/pathology*
;
Capsules
;
DNA, Viral/analysis*
;
Drugs, Chinese Herbal/therapeutic use*
;
Feces/virology*
;
Female
;
Humans
;
Length of Stay
;
Lymphocyte Count
;
Male
;
Medicine, Chinese Traditional/methods*
;
Middle Aged
;
Retrospective Studies
;
SARS-CoV-2/genetics*
;
Severity of Illness Index
;
Treatment Outcome
;
Young Adult
4.High-throughput screening identifies established drugs as SARS-CoV-2 PLpro inhibitors.
Yao ZHAO ; Xiaoyu DU ; Yinkai DUAN ; Xiaoyan PAN ; Yifang SUN ; Tian YOU ; Lin HAN ; Zhenming JIN ; Weijuan SHANG ; Jing YU ; Hangtian GUO ; Qianying LIU ; Yan WU ; Chao PENG ; Jun WANG ; Chenghao ZHU ; Xiuna YANG ; Kailin YANG ; Ying LEI ; Luke W GUDDAT ; Wenqing XU ; Gengfu XIAO ; Lei SUN ; Leike ZHANG ; Zihe RAO ; Haitao YANG
Protein & Cell 2021;12(11):877-888
A new coronavirus (SARS-CoV-2) has been identified as the etiologic agent for the COVID-19 outbreak. Currently, effective treatment options remain very limited for this disease; therefore, there is an urgent need to identify new anti-COVID-19 agents. In this study, we screened over 6,000 compounds that included approved drugs, drug candidates in clinical trials, and pharmacologically active compounds to identify leads that target the SARS-CoV-2 papain-like protease (PLpro). Together with main protease (M
Antiviral Agents/therapeutic use*
;
Binding Sites
;
COVID-19/virology*
;
Coronavirus Papain-Like Proteases/metabolism*
;
Crystallography, X-Ray
;
Drug Evaluation, Preclinical
;
Drug Repositioning
;
High-Throughput Screening Assays/methods*
;
Humans
;
Imidazoles/therapeutic use*
;
Inhibitory Concentration 50
;
Molecular Dynamics Simulation
;
Mutagenesis, Site-Directed
;
Naphthoquinones/therapeutic use*
;
Protease Inhibitors/therapeutic use*
;
Protein Structure, Tertiary
;
Recombinant Proteins/isolation & purification*
;
SARS-CoV-2/isolation & purification*
5.Analysis of an improved workflow of endoscope reprocessing for bedside endoscopic diagnosis and treatment on COVID-19 patients.
Qing GU ; Hua-Fen WANG ; Ying FANG ; Ye LU ; Zhe SHEN ; Yan WANG ; Xin WU ; Li CEN ; Yi-Shu CHEN
Journal of Zhejiang University. Science. B 2020;21(5):416-422
Severe cases infected with the coronavirus disease 2019 (COVID-19), named by the World Health Organization (WHO) on Feb. 11, 2020, tend to present a hypercatabolic state because of severe systemic consumption, and are susceptible to stress ulcers and even life-threatening gastrointestinal bleeding. Endoscopic diagnosis and treatment constitute an irreplaceable part in the handling of severe COVID-19 cases. Endoscopes, as reusable precision instruments with complicated structures, require more techniques than other medical devices in cleaning, disinfection, sterilization, and other reprocessing procedures. From 2016 to 2019, health care-acquired infection caused by improper endoscope reprocessing has always been among the top 5 on the list of top 10 health technology hazards issued by the Emergency Care Research Institute. Considering the highly infective nature of COVID-19 and the potential aerosol contamination therefrom, it is of pivotal significance to ensure that endoscopes are strictly reprocessed between uses. In accordance with the national standard "Regulation for Cleaning and Disinfection Technique of Flexible Endoscope (WS507-2016)," we improved the workflow of endoscope reprocessing including the selection of chemicals in an effort to ensure quality control throughout the clinical management towards COVID-19 patients. Based on the experience we attained from the 12 severe COVID-19 cases in our hospital who underwent endoscopy 23 times in total, the article provides an improved version of endoscopic reprocessing guidelines for bedside endoscopic diagnosis and treatment on COVID-19 patients for reference.
Adult
;
Aged
;
Aged, 80 and over
;
Betacoronavirus
;
China
;
Coronavirus Infections
;
diagnosis
;
therapy
;
Cross Infection
;
prevention & control
;
Disinfection
;
methods
;
Endoscopes
;
virology
;
Equipment Contamination
;
prevention & control
;
Female
;
Humans
;
Male
;
Middle Aged
;
Pandemics
;
Peracetic Acid
;
Personal Protective Equipment
;
Pneumonia, Viral
;
diagnosis
;
therapy
;
Sterilization
;
methods
;
Workflow
6.Progress in engineering application of human adenovirus.
Yang ZHAO ; Qiwei ZHANG ; Xueshan XIA
Chinese Journal of Biotechnology 2020;36(7):1269-1276
Human adenoviruses are widespread causative agent that induces respiratory diseases, epidemic keratoconjunctivitis and other related diseases. Adenoviruses are commonly used in experimental and clinical areas. It is one of the most commonly used virus vectors in gene therapy, and it has attracted a lot of attention and has a high research potential in tumor gene therapy and virus oncolytic. Here, we summarize the biological characteristics, epidemiology and current application of adenovirus, in order to provide reference for engineering application of adenovirus.
Adenovirus Infections, Human
;
epidemiology
;
virology
;
Adenoviruses, Human
;
genetics
;
Genetic Engineering
;
methods
;
trends
;
Genetic Vectors
;
Humans
;
Oncolytic Virotherapy
;
trends
;
Oncolytic Viruses
;
genetics
;
Virus Replication
7.Optimization of a fluorescent qPCR detection for RNA of SARS-CoV-2.
Xuelong LI ; Junhua LIU ; Qianyang LIU ; Lin YU ; Shanshan WU ; Xiushan YIN
Chinese Journal of Biotechnology 2020;36(4):732-739
We optimized a fluorescent quantitative polymerase chain reaction (qPCR) assay system for rapid and real time detection of SARS-CoV-2 RNA. The results show that the lowest dilution of RNA samples used for the detection of SARS-CoV-2 RNA could reach 1/10 000 (the initial value is set as 10 ng/μL). Moreover, the cycle threshold (Ct) for samples of clinically diagnosed COVID-19 was lower than 35 or 40. The sensitivity of this method was satisfactory. The results were consistent with those of the COVID-19 detection kit on the market under the same conditions, but the number of cycles required was shortened by about 2. Therefore, the optimized assay developed in this study can be used in screening and early clinical diagnosis. Our work provides a tool to facilitate rapid clinical diagnosis of COVID-19.
Betacoronavirus
;
genetics
;
isolation & purification
;
Coronavirus Infections
;
diagnosis
;
virology
;
Early Diagnosis
;
Humans
;
Pandemics
;
Pneumonia, Viral
;
diagnosis
;
virology
;
Polymerase Chain Reaction
;
methods
;
standards
;
RNA, Viral
;
analysis
;
genetics
;
Sensitivity and Specificity
;
Time Factors
10.Traditional Chinese medicine network pharmacology study on exploring the mechanism of Xuebijing Injection in the treatment of coronavirus disease 2019.
Yan XING ; Ying-Rong HUA ; Jing SHANG ; Wei-Hong GE ; Jun LIAO
Chinese Journal of Natural Medicines (English Ed.) 2020;18(12):941-951
As a representative drug for the treatment of severe community-acquired pneumonia and sepsis, Xuebijing (XBJ) injection is also one of the recommended drugs for the prevention and treatment of coronavirus disease 2019 (COVID-19), but its treatment mechanism for COVID-19 is still unclear. Therefore, this study aims to explore the potential mechanism of XBJ injection in the treatment of COVID-19 employing network pharmacology and molecular docking methods. The corresponding target genes of 45 main active ingredients in XBJ injection and COVID-19 were obtained by using multiple database retrieval and literature mining. 102 overlapping targets of them were screened as the core targets for analysis. Then built the PPI network, TCM-compound-target-disease, and disease-target-pathway networks with the help of Cytoscape 3.6.1 software. After that, utilized DAVID to perform gene ontology (GO) function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis to predict the action mechanism of overlapping targets. Finally, by applying molecular docking technology, all compounds were docked with COVID-19 3 CL protease(3CLpro), spike protein (S protein), and angiotensin-converting enzyme II (ACE2). The results indicated that quercetin, luteolin, apigenin and other compounds in XBJ injection could affect TNF, MAPK1, IL6 and other overlapping targets. Meanwhile, anhydrosafflor yellow B (AHSYB), salvianolic acid B (SAB), and rutin could combine with COVID-19 crucial proteins, and then played the role of anti-inflammatory, antiviral and immune response to treat COVID-19. This study revealed the multiple active components, multiple targets, and multiple pathways of XBJ injection in the treatment of COVID-19, which provided a new perspective for the study of the mechanism of traditional Chinese medicine (TCM) in the treatment of COVID-19.
Angiotensin-Converting Enzyme 2/metabolism*
;
Biological Availability
;
COVID-19/virology*
;
Coronavirus 3C Proteases/metabolism*
;
Drugs, Chinese Herbal/therapeutic use*
;
Humans
;
Medicine, Chinese Traditional/methods*
;
Molecular Docking Simulation/methods*
;
Protein Interaction Mapping/methods*
;
SARS-CoV-2/physiology*
;
Signal Transduction/drug effects*
;
Spike Glycoprotein, Coronavirus/metabolism*

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