This study aims to establish an antibody detection method for porcine deltacoronavirus (PDCoV). The recombinant proteins PDCoV-N1 and PDCoV-N2 were expressed via the prokaryotic plasmid pColdII harboring the N gene sequence of the PDCoV strain CH/XJYN/2016. The reactivity and specificity of PDCoV-N1 and PDCoV-N2 with anti-PEDV sera were analyzed after the recombinant proteins were analyzed by SDS-PAGE and purified by the Ni-NTA Superflow Cartridge. Meanwhile, Western blotting and indirect immunofluorescence assay were carried out separately to validate the recombinant proteins PDCoV-N1 and PDCoV-N2. Finally, we established an indirect ELISA method based on the recombinant protein PDCoV-N2 after optimizing the conditions and tested the sensitivity, specificity, and reproducibility of the method. Then, the established method was employed to examine 102 clinical serum samples. The recombinant protein PDCoV-N2 showed low cross-reactivity with anti-PEDV sera. The optimal conditions of the indirect ELISA method based on PDCoV-N2 were as follows: the antigen coating concentration of 1.25 μg/mL and coating at 37 ℃ for 1 h; blocking by BSA overnight at 4 ℃; serum sample dilution at 1:50 and incubation at 37 ℃ for 1 h; secondary antibody dilution at 1:80 000 and incubation at 37 ℃ for 1 h; color development with TMB chromogenic solution at 37 ℃ for 10 min. The S/P value ≥ 0.45, ≤0.38, and between 0.45 and 0.38 indicated that the test sample was positive, negative, and suspicious, respectively. The testing results of the antisera against porcine epidemic diarrhea virus (PEDV), porcine circovirus 2 (PCV2), transmissible gastroenteritis virus (TGEV), foot-and-mouth disease virus (FMDV), and African swine fever virus (ASFV) showed that the S/P values were all less than 0.38. The testing results of the 800-fold diluted anti-PDCoV sera were still positive. The results of the inter- and intra-batch tests showed that the coefficients of variation of this method were less than 10%. Clinical serum sample test results showed the coincidence rate between this method and neutralization test was 94.12%. In this study, an ELISA method for the detection of anti-PDCoV antibodies was successfully established based on the truncated N protein of PDCoV. This method is sensitive, specific, stable, and reproducible, serving as a new method for the clinical diagnosis of PDCoV.
Animals ; Enzyme-Linked Immunosorbent Assay/methods* ; Swine ; Antibodies, Viral/blood* ; Recombinant Proteins/genetics* ; Deltacoronavirus/isolation & purification* ; Coronavirus Infections/virology* ; Swine Diseases/diagnosis* ; Coronavirus Nucleocapsid Proteins ; Sensitivity and Specificity

Fever is an increase in body temperature beyond the normal range, acting as a protective inflammatory mechanism. This article summarizes diseases with fever encountered in dental clinics, including what is known about pyrexia in coronavirus infection, and further proposes a "six steps in one" identification and analysis strategy to guide the clinical work of stomatology.
Humans ; Dental Clinics ; Fever/diagnosis* ; Coronavirus Infections

In the past, coronavirus caused two serious human-to-human pandemics in the world, including severe acute respiratory syndrome （SARS） and Middle East respiratory syndrome （MERS）. In late 2019, coronavirus disease 2019 （COVID-19） caused another major global public health event. Due to the strong infectivity of novel coronavirus, it is difficult to carry out the autopsy of related death cases widely. This paper reviews the previous status of the pathogen detection related to the autopsy of coronavirus infection diseases, and introduces the ongoing detection methods of novel coronavirus in clinical practice, in order to provide reference for the pathogen detection and study related to autopsy of COVID-19.
Autopsy ; COVID-19 ; Communicable Diseases ; Coronavirus Infections/diagnosis* ; Humans ; Middle East Respiratory Syndrome Coronavirus ; SARS-CoV-2

Betacoronavirus ; Coronavirus Infections ; epidemiology ; Humans ; Melanoma ; diagnosis ; therapy ; Pandemics ; Pneumonia, Viral ; epidemiology ; Skin Neoplasms ; diagnosis ; therapy

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

As of Apr. 22, 2020, the World Health Organization (2020) has reported over 2.4 million confirmed coronavirus disease 2019 (COVID-19) patients and 169 151 deaths. Recent articles have uncovered genomic characteristics and clinical features of COVID-19 (Chan et al., 2020; Chang et al., 2020; Guan et al., 2020; Zhu et al., 2020), while our understanding of COVID-19 is still limited. As suggested by guidelines promoted by the General Office of National Health Commission of the People's Republic of China (2020) (from Versions 1 to 6), discharged standards for COVID-19 were still dependent on viral real-time polymerase chain reaction (RT-PCR) tests of respiratory specimens, showing that recovered COVID-19 patients with twice negative RT-PCR could meet discharge criteria. Here, we examined two cases in which nucleic acid test results were inconsistent with clinical and radiological findings, leading to suboptimal care.
Adult ; Betacoronavirus ; China ; Clinical Laboratory Techniques ; Coronavirus Infections ; diagnosis ; transmission ; Female ; Humans ; Male ; Middle Aged ; Pandemics ; Patient Discharge ; Pneumonia, Viral ; diagnosis ; transmission ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Sputum ; virology

The recently emerged novel coronavirus pneumonia, named the coronavirus disease 2019 (COVID-19), shares several clinical characteristics with severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS), and spread rapidly throughout China in December of 2019 (Huang et al., 2020). The pathogen 2019 novel coronavirus (2019-nCoV) is now named SARS coronavirus 2 (SARS-CoV-2) and is highly infectious. As of Apr. 9, 2020, over 80 000 confirmed cases had been reported, with an estimated mortality rate of 4.0% (Chinese Center for Disease Control and Prevention, 2020). Person-to-person transmission and familial clustering have been reported (Chan et al., 2020; Nishiura et al., 2020; Phan et al., 2020). However, there is no evidence of fetal intrauterine infection in pregnant women who have been infected with SARS-CoV-2 in their third trimester (Chen et al., 2020). It is unclear whether breastfeeding transmits the virus from previously infected and recovered mothers to their babies. Here we report the clinical course of a pregnant woman with COVID-19. In order to determine whether SARS-CoV-2 can be transmitted to newborns through breastfeeding, we measured viral RNA in the patient's breastmilk samples at different time points after delivery.
Adult ; Betacoronavirus ; Breast Feeding ; China ; Coronavirus Infections ; diagnosis ; Female ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Milk, Human ; virology ; Pandemics ; Pneumonia, Viral ; diagnosis ; Pregnancy ; Pregnancy Complications, Infectious ; virology ; RNA, Viral ; isolation & purification

Since the global outbreak of severe acute respiratory syndrome (SARS) in 2003, China has gradually built a robust prevention and control system for sudden infectious diseases. All large hospitals have a fever clinic that isolates patients with all kinds of acute communicable diseases as the first line of medical defense. The emergency department, as the second line of medical defense in hospitals, is constantly shouldering the heavy responsibility of screening communicable diseases while also treating all kinds of other non-communicable acute and critical diseases (Zhang et al., 2012; Zhu et al., 2015; Wang et al., 2017; Feng et al., 2018; Lu, 2018; Xu and Lu, 2019). An outbreak of pneumonia of unknown etiology that began in Wuhan city (China) has spread rapidly in China since December 2019 (Huang et al., 2020; WHO, 2020; Zhu et al., 2020). In February 2020, the National Health Commission of China named the disease a novel coronavirus pneumonia (NCP); then, it was formally named the coronavirus disease 2019 (COVID-19) by the World Health Organization (WHO) on Feb. 11, 2020. The Coronavirus Study Group of the International Committee on Taxonomy of Viruses designated this causative virus as SARS coronavirus 2 (SARS-CoV-2). SARS-CoV-2 belongs to the β coronavirus genus, and its pathogenic mechanism has not been clarified, which requires further study. To better understand the clinical characteristics of COVID-19 and more effectively prevent and control this disease, we retrospectively analyzed four representative cases of COVID-19 that had recently been screened and diagnosed in our emergency department.
Adult ; Betacoronavirus ; China ; epidemiology ; Coronavirus Infections ; diagnosis ; Emergency Service, Hospital ; Female ; Humans ; Lung ; diagnostic imaging ; Male ; Middle Aged ; Pandemics ; Patient Isolation ; Pneumonia, Viral ; diagnosis ; Tomography, X-Ray Computed

Pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection emerged in Wuhan City, Hubei Province, China in December 2019. By Feb. 11, 2020, the World Health Organization (WHO) officially named the disease resulting from infection with SARS-CoV-2 as coronavirus disease 2019 (COVID-19). COVID-19 represents a spectrum of clinical manifestations that typically include fever, dry cough, and fatigue, often with pulmonary involvement. SARS-CoV-2 is highly contagious and most individuals within the population at large are susceptible to infection. Wild animal hosts and infected patients are currently the main sources of disease which is transmitted via respiratory droplets and direct contact. Since the outbreak, the Chinese government and scientific community have acted rapidly to identify the causative agent and promptly shared the viral gene sequence, and have carried out measures to contain the epidemic. Meanwhile, recent research has revealed critical aspects of SARS-CoV-2 biology and disease pathogenesis; other studies have focused on epidemiology, clinical features, diagnosis, management, as well as drug and vaccine development. This review aims to summarize the latest research findings and to provide expert consensus. We will also share ongoing efforts and experience in China, which may provide insight on how to contain the epidemic and improve our understanding of this emerging infectious disease, together with updated guidance for prevention, control, and critical management of this pandemic.
Amino Acid Motifs ; Animals ; Antiviral Agents ; Betacoronavirus ; genetics ; China ; epidemiology ; Communicable Disease Control ; methods ; Coronavirus Infections ; diagnosis ; epidemiology ; physiopathology ; prevention & control ; therapy ; Humans ; Immunization, Passive ; Medicine, Chinese Traditional ; Pandemics ; Pneumonia, Viral ; diagnosis ; epidemiology ; physiopathology ; therapy ; Protein Domains ; Spike Glycoprotein, Coronavirus ; chemistry ; Viral Vaccines

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