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

Adult ; Aged ; Betacoronavirus ; COVID-19 ; Coronavirus Infections/diagnosis* ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Pandemics ; Pneumonia/diagnosis* ; Pneumonia, Viral/diagnosis* ; Retrospective Studies ; SARS-CoV-2

OBJECTIVE: To evaluate the application of three-in-one intelligent screening in outpatient pre-inspection in children's hospital. METHODS: We randomly enrolled 100 children pre-screened by traditional method in the outpatient department of Children's Hospital of Zhejiang University from February 6th to 16th, 2020, and another 100 children by the intelligent three-in-one mode from February 17th to 27th, 2020. The traditional triage was conducted by nurses based on face-to-face, one-by-one interview of the epidemiological history and consultation department, and the temperature was measured before manual triage. The intelligent three-in-one model combined online rapid pre-inspection and triage, on-site manual confirmation, as well as synchronized online health education system. For on-line registered patients, the system automatically sent the COVID-19 epidemiological pre-screening triage questionnaire one hour before the appointment, requiring parents to complete and submit online before arriving at the hospital. The on-site registered patients were controlled at 100 m away from the hospital entrance. The nurses guided the parents to scan the QR code and fill in the COVID-19 epidemiological pre-examination triage questionnaire. At the entrance of the hospital, the nurse checked the guidance sheet and took the temperature again. The children with red guidance sheet were checked again and confirmed by pre-examination nurses, and accompanied to the isolation clinic through COVID-19 patients-only entrance. The children with yellow guidance sheet were guided to fever clinic. The children with green guidance sheet could go with their parents to the designated area, and then went to the corresponding consultation area. Health education was carried out throughout the treatment, and the system automatically posted the corresponding outpatient instructions and education courses. Parents would read the courses on their mobile phones and counsel online. The time of pre-examination and the coincidence rate of triage were compared between the two groups. RESULTS: The three-in-one intelligent pre-inspection mode took an average of (25.6±8.0) s for each child, which was significantly shorter than the traditional pre-inspection mode (74.8±36.4) s ( CONCLUSIONS The three-in-one intelligent pre-inspection model can effectively shorten the patient pre-check time, with similar triage coincidence rate to traditional model.
Adult ; Betacoronavirus ; COVID-19 ; Child ; Coronavirus Infections/diagnosis* ; Humans ; Internet ; Outpatient Clinics, Hospital ; Pandemics ; Pneumonia, Viral/diagnosis* ; SARS-CoV-2 ; Surveys and Questionnaires ; Time ; Triage/standards*

OBJECTIVE: To evaluate the effect of standardized health education on the sputum specimen collection rate for nucleic acid detection of coronavirus disease 2019 (COVID-19). METHODS: Two hundred and twenty-seven patients in fever clinics and isolation wards of Sir Run Run Shaw Hospital of Zhejiang University and 307 migrant workers returning to 5 enterprises in Shanghai from February 3 to March 14, 2020 were enrolled in the study. Through clarifying the procedures of collecting sputum specimens, making graphic/video health education materials, standardizing the contents and methods of health education, we conducted education to the subjects. The subject expectorated spontaneously or with medical assistance. For patients, the number of sampling attempts and sputum acquisition times were documented before and after the implementation of the standardized expectoration method; for the returning migrant employees in the enterprises, only the number of collected samples after the implementation of the standardized expectoration method were recorded. RESULTS: A total of 378 sputum samples were collected from 227 patients. The sputum sampling rates before and after the implementation of health education were 40.9%and 58.4%, respectively ( CONCLUSIONS The education for standardized sputum sample collection method can effectively increase the sputum collection rate.
Betacoronavirus/genetics* ; COVID-19 ; China ; Coronavirus Infections/diagnosis* ; Efficiency ; Humans ; Nucleic Acid Amplification Techniques/methods* ; Pandemics ; Pneumonia, Viral/diagnosis* ; SARS-CoV-2 ; Specimen Handling/methods* ; Sputum ; Time Factors

OBJECTIVE: Coronavirus disease 2019 (COVID-19) has raised concern around the world as an epidemic or pandemic. As data on COVID-19 has grown, it has become clear that older adults have a disproportionately high rate of death from COVID-19. This study describes the early clinical characteristics of COVID-19 in patients with more than 80 years of age. METHODS: Epidemiological, clinical, laboratory, radiological, and treatment data from 17 patients diagnosed with COVID-19 between January 20 and February 20, 2020 were collected and analyzed retrospectively. Treatment outcomes among subgroups of patients with non-severe and severe symptoms of COVID-19 were compared. RESULTS: Of the 17 hospitalized patients with COVID-19, the median age was 88.0 years (interquartile range, 86.6-90.0 years; range, 80.0-100.0 years) and 12 (70.6%) were men. The age distribution of patients was not significantly different between non-severe group and severe group. All patients had chronic pre-existing conditions. Hypertension and cardiovascular diseases were the most common chronic conditions in both subgroups. The most common symptoms at the onset of COVID-19 were fever (n = 13; 76.5%), fatigue (n = 11; 64.7%), and cough (n = 5; 29.4%). Lymphopenia was observed in all patients, and lymphopenia was significantly more severe in the severe group than that in non-severe group (0.4 × 10/L vs 1.2 × 10/L, P = 0.014). The level of serum creatinine was higher in the severe group than in the non-severe group (99.0 μmol/L vs 62.5 μmol/L, P = 0.038). The most common features of chest computed tomography images were nodular foci in 10 (58.8%) patients and pleural thickening in 7 (41.2%) patients. All patients received antiviral therapy, while some patients also received intravenous antibiotics therapy (76.5%), Chinese medicinal preparation therapy (Lianhuaqingwen capsule, 64.7%), corticosteroids (35.3%) or immunoglobin (29.4%). Eight patients (47.1%) were transferred to the intensive care unit because of complications. Ten patients (58.8%) received intranasal oxygen, while 3 (17.6%) received non-invasive mechanical ventilation, and 4 (23.5%) received high-flow oxygen. As of June 20, 7 (41.2%) patients had been discharged and 10 (58.8% of this cohort, 77.8% of severe patients) had died. CONCLUSION The mortality of patients aged 80 years and older with severe COVID-19 symptoms was high. Lymphopenia was a characteristic laboratory result in these patients, and the severity of lymphopenia was indicative of the severity of COVID-19. However, the majority of patients with COVID-19 in this age cohort had atypical symptoms, and early diagnosis depends on prompt use of a viral nucleic acid test.
Age Factors ; Aged, 80 and over ; Antiviral Agents ; therapeutic use ; China ; epidemiology ; Clinical Laboratory Techniques ; Coronavirus Infections ; diagnosis ; epidemiology ; pathology ; Female ; Humans ; Lung ; diagnostic imaging ; pathology ; Male ; Pandemics ; Pneumonia, Viral ; diagnosis ; epidemiology ; pathology ; Retrospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

Since the outbreak of novel coronavirus pneumonia (coronavirus disease 2019, COVID-19), it has rapidly spread to 187 countries, causing serious harm to the health of people and a huge social burden. However, currently, drugs specifically approved for clinical use are not available, except for vaccines against COVID-19 that are being evaluated. Traditional Chinese medicine (TCM) is capable of performing syndrome differentiation and treatment according to the clinical manifestations of patients, and has a better ability of epidemic prevention and control. The authors comprehensively analyzed the etiology and pathogenesis of COVID-19 based on the theory of TCM, and discussed its syndrome differentiation, treatment and prevention measures so as to provide strategies and reference for the prevention and treatment with TCM.
Betacoronavirus ; Coronavirus Infections ; diagnosis ; etiology ; prevention & control ; therapy ; Humans ; Medicine, Chinese Traditional ; Pandemics ; prevention & control ; Pneumonia, Viral ; diagnosis ; etiology ; prevention & control ; therapy

OBJECTIVE: To early differentiate between coronavirus disease 2019 (COVID-19) and adult mycoplasma pneumonia with chest CT scan. METHODS: Twenty-six patients with COVID-19 and 21 patients with adult mycoplasma pneumonia confirmed with RT-PCR test were enrolled from Zibo First Hospital and Lanshan People's Hospital during December 1st 2019 and March 14th 2020. The early chest CT manifestations were analyzed and compared between the two groups. RESULTS: The interstitial changes with ground glass density shadow (GGO) were similar in two groups during first chest CT examination (>0.05). There were more lung lobes involved on the first chest CT in COVID-19 patients, which were mostly distributed in the dorsal outer zone (23/26, 88.5%), and nearly half of them (12/26, 46.2%) were accompanied by crazy-paving sign; while the lesions in adult mycoplasma pneumonia patients were mostly distributed along the bronchi, and the bronchial wall was thickened (19/21, 90.5%), accompanied with tree buds / fog signs (19/21, 90.5%). The above CT signs were significantly different between the two kinds of pneumonia (all <0.01). COVID-19 had a longer course compared with mycoplasma pneumonia, the disease peaks of COVID-19 patients was on day (10.5±3.8), while the disease on CT was almost absorbed on day (7.9±2.2) in adult mycoplasma pneumonia. The length of hospital stay in COVID-19 patients was significantly longer than that of mycoplasma pneumonia patients [(19.5±4.3) d vs (7.9±2.2) d, <0.01]. CONCLUSIONS The lesions of adult mycoplasma pneumonia are mostly distributed along the bronchi with tree buds/fog signs, while the lesions of COVID-19 are mainly distributed in the dorsal outer zone accompanied by crazy-paving sign, which can early distinguish two diseases.
Adult ; Betacoronavirus ; Clinical Laboratory Techniques ; standards ; Coronavirus Infections ; diagnosis ; diagnostic imaging ; Diagnosis, Differential ; Humans ; Lung ; diagnostic imaging ; Pandemics ; Pneumonia, Mycoplasma ; diagnostic imaging ; Pneumonia, Viral ; diagnostic imaging ; Tomography, X-Ray Computed

OBJECTIVE: To identify the key biochemical indicators that affect the clinical type and outcomes of COVID-19 patients and explore the application of neutrophil/lymphocyte ratio (NLR) in COVID-19. METHODS: Ninety-three patients with confirmed diagnosis of COVID-19 admitted in Ezhou Central Hospital from February to April in 2020 were analyzed. Among them, 43 patients were selected from Intensive Care Unit (ICU) with the diagnosis of critical type of COVID-19, and 50 cases of common type were selected from the Department of Respiratory Medicine. The baseline data, blood routine test and biochemical indexes of the patients were collected on the first day of admission. NLRs of the patients were calculated, and COX survival analysis according to the NLR 4-category method was performed. The patients' outcomes were analyzed with receiver operating curves (ROCs). The patients were divided into two groups according to NLR cutoff value for comparison of the biochemical indexes. Based on the patients' outcomes, NLR cutoff value classification and clinical classification, multiple binary logistics regression was performed to screen the key variables and explore their significance in COVID-19. RESULTS: The NLR four-category method was not applicable for prognostic evaluation of the patients. The cut-off value of NLR for predict the prognosis of COVID-19 was 11.26, with a sensitivity of 0.903 and a specificity of 0.839; the laboratory indicators of the patients with NLR < 11.26 were similar to those in patients of the common type; the indicators were also similar between patients with NLR≥11.26 and those with critical type COVID-19. NLR, WBC, NEUT, PCT, DD, BUN, TNI, BNP, and LDH had significant effects on the clinical classification and outcome of the patients ( < 0.05); Cr, Ca, PH, and Lac had greater impact on the outcome of the patients ( < 0.05), while Na, PCO had greater impact on the clinical classification of the patients ( < 0.05). CONCLUSIONS NLR can be used as an important reference for clinical classification, prognostic assessment, and biochemical abnormalities of COVID-19. Patients of critical type more frequently have bacterial infection with more serious inflammatory reactions, severer heart, lung and kidney damages, and much higher levels of DD and LDH than those of the common type. NLR, NEUT, DD, TNI, BNP, LDH, Ca, PCT, PH, and Lac have obvious influence on the prognosis of COVID-19 and should be observed dynamically.
Betacoronavirus ; Blood Cell Count ; standards ; Coronavirus Infections ; blood ; diagnosis ; physiopathology ; Humans ; Lymphocytes ; cytology ; Neutrophils ; cytology ; Pandemics ; Pneumonia, Viral ; blood ; diagnosis ; physiopathology ; Prognosis ; ROC Curve ; Retrospective Studies ; Severity of Illness Index