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1.
Cureus ; 15(2): e35534, 2023 Feb.
Article in English | MEDLINE | ID: covidwho-2257864

ABSTRACT

Introduction California State Bill 1152 (SB1152) mandated all non-state-operated hospitals meet specific criteria when discharging patients identified as experiencing homelessness. Little is known about SB1152's effect on hospitals or compliance statewide. We studied the implementation of SB1152 in our emergency department (ED). Methods We analyzed our suburban academic ED's institutional electronic medical record for one year before (July 1, 2018-June 20, 2019) and one year after (July 1, 2019-June 30, 2020) implementation of SB1152. We identified individuals by lack of address during registration, International Classification of Diseases, Tenth Revision (ICD-10) code of homelessness, and/or the presence of an SB1152 discharge checklist. Demographics, clinical information, and repeat visit data were collected. Results ED volumes were constant during the pre- and post-SB1152 periods (approximately 75,000 annually); however, ED visits by people experiencing homelessness more than doubled (630 (0.8%) to 1530 (2.1%) in the pre- and post-implementation periods. Age and sex distributions were similar with approximately 80% of patients aged 31-65 years and less than 1% under 18. Visits by females comprised less than 30% of the population. Visits by people of the White race decreased from 50% to 40% pre- and post-SB1152. Visits by people of the Black, Asian, and Hispanic races experiencing homelessness increased by 18% to 25%, 1% to 4%, and 19% to 21%, respectively. Acuity was unchanged with 50% of visits classified as "urgent." Discharges increased from 73% to 81% and admissions halved from 18% to 9%. Visits by patients with only one ED visit decreased (28% to 22%); those with four or more visits increased (46% to 56%). The most common primary diagnoses pre- and post-SB1162 were alcohol use (6.8% and 9.3%, respectively), chest pain (3.3% and 4.5%, respectively), convulsions (3.0%, and 2.46%, respectively), and limb pain (2.3% and 2.3%, respectively). The primary diagnosis of suicidal ideation doubled from the pre- to post-implementation periods (1.3% to 2.2%, respectively). Checklists were completed for 92% of identified patients discharged from the ED. Conclusion Implementation of SB1152 in our ED resulted in identifying an increased number of persons experiencing homelessness. We identified opportunities for further improvement since pediatric patients were missed. Further analysis is warranted, especially with the coronavirus disease 2019 (COVID-19) pandemic, which has significantly affected healthcare-seeking behavior in EDs.

2.
ANZ J Surg ; 93(4): 932-938, 2023 04.
Article in English | MEDLINE | ID: covidwho-2213471

ABSTRACT

BACKGROUND: Malignant polyps represent the early development of colorectal adenocarcinoma. During 2020, there was widescale rationing of health-care resources in response to the COVID-19 pandemic. In particular there was deferral of some colonoscopy procedures required for timely malignant polyp detection. This study sought to assess how these deferrals affected the diagnosis of malignant polyps. METHODS: A population wide analysis was performed of 2079 malignant polyps, diagnosed in Queensland, Australia from 2011 to 2020. A regression analysis, with 95% prediction intervals, was produced to determine whether there was a significant impact on the number of malignant polyps diagnosed in 2020 compared to previous years. Univariate statistical analysis of patient, procedural, and pathological variables was also performed. RESULTS: In 2020 there were 211 malignant polyps diagnosed, which was significantly lower than was predicted by the univariate regression analysis (r2  = 0.85, 95% prediction interval: 255.07-323.91, P < 0.001). These malignant polyps were less likely to be diagnosed in a private setting (P < 0.001), and exhibited significantly less depth of submucosal invasion (P = 0.017). There was no significant difference in the management strategy (polypectomy, resection or trans-anal resection) between 2011 and 2019 and 2020. CONCLUSION: Because of the significant decrease in the number of malignant polyps, and the natural history of the disease, it is expected that there will be an increase in more advanced colorectal adenocarcinomas presenting in 2021 and beyond. This has implications for healthcare resources, particularly in light of the ongoing strain on health departments as a result of the COVID-19 pandemic.


Subject(s)
Adenocarcinoma , COVID-19 , Colonic Polyps , Colorectal Neoplasms , Humans , Colonic Polyps/epidemiology , Colonic Polyps/surgery , Colonic Polyps/pathology , Pandemics , COVID-19/epidemiology , Colonoscopy , Colorectal Neoplasms/pathology , Adenocarcinoma/surgery
3.
J Am Coll Emerg Physicians Open ; 3(6): e12867, 2022 Dec.
Article in English | MEDLINE | ID: covidwho-2172887

ABSTRACT

Objectives: Here we report the clinical performance of COVID-19 curbside screening with triage to a drive-through care pathway versus main emergency department (ED) care for ambulatory COVID-19 testing during a pandemic. Patients were evaluated from cars to prevent the demand for testing from spreading COVID-19 within the hospital. Methods: We examined the effectiveness of curbside screening to identify patients who would be tested during evaluation, patient flow from screening to care team evaluation and testing, and safety of drive-through care as 7-day ED revisits and 14-day hospital admissions. We also compared main ED efficiency versus drive-through care using ED length of stay (EDLOS). Standardized mean differences (SMD) >0.20 identify statistical significance. Results: Of 5931 ED patients seen, 2788 (47.0%) were walk-in patients. Of these patients, 1111 (39.8%) screened positive for potential COVID symptoms, of whom 708 (63.7%) were triaged to drive-through care (with 96.3% tested), and 403 (36.3%) triaged to the main ED (with 90.5% tested). The 1677 (60.2%) patients who screened negative were seen in the main ED, with 440 (26.2%) tested. Curbside screening sensitivity and specificity for predicting who ultimately received testing were 70.3% and 94.5%. Compared to the main ED, drive-through patients had fewer 7-day ED revisits (3.8% vs 12.5%, SMD = 0.321), fewer 14-day hospital readmissions (4.5% vs 15.6%, SMD = 0.37), and shorter EDLOS (0.56 vs 5.12 hours, SMD = 1.48). Conclusion: Curbside screening had high sensitivity, permitting early respiratory isolation precautions for most patients tested. Low ED revisit, hospital readmissions, and EDLOS suggest drive-through care, with appropriate screening, is safe and efficient for future respiratory illness pandemics.

4.
J Mol Diagn ; 24(4): 320-336, 2022 04.
Article in English | MEDLINE | ID: covidwho-1895234

ABSTRACT

Previous studies have described reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for the rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal/oropharyngeal swab and saliva samples. This multisite clinical evaluation describes the validation of an improved sample preparation method for extraction-free RT-LAMP and reports clinical performance of four RT-LAMP assay formats for SARS-CoV-2 detection. Direct RT-LAMP was performed on 559 swabs and 86,760 saliva samples and RNA RT-LAMP on extracted RNA from 12,619 swabs and 12,521 saliva samples from asymptomatic and symptomatic individuals across health care and community settings. For direct RT-LAMP, overall diagnostic sensitivity (DSe) was 70.35% (95% CI, 63.48%-76.60%) on swabs and 84.62% (95% CI, 79.50%-88.88%) on saliva, with diagnostic specificity of 100% (95% CI, 98.98%-100.00%) on swabs and 100% (95% CI, 99.72%-100.00%) on saliva, compared with quantitative RT-PCR (RT-qPCR); analyzing samples with RT-qPCR ORF1ab CT values of ≤25 and ≤33, DSe values were 100% (95% CI, 96.34%-100%) and 77.78% (95% CI, 70.99%-83.62%) for swabs, and 99.01% (95% CI, 94.61%-99.97%) and 87.61% (95% CI, 82.69%-91.54%) for saliva, respectively. For RNA RT-LAMP, overall DSe and diagnostic specificity were 96.06% (95% CI, 92.88%-98.12%) and 99.99% (95% CI, 99.95%-100%) for swabs, and 80.65% (95% CI, 73.54%-86.54%) and 99.99% (95% CI, 99.95%-100%) for saliva, respectively. These findings demonstrate that RT-LAMP is applicable to a variety of use cases, including frequent, interval-based direct RT-LAMP of saliva from asymptomatic individuals who may otherwise be missed using symptomatic testing alone.


Subject(s)
COVID-19 , SARS-CoV-2 , COVID-19/diagnosis , COVID-19 Testing , Humans , Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , RNA, Viral/analysis , RNA, Viral/genetics , SARS-CoV-2/genetics , Saliva , Sensitivity and Specificity
5.
Int J Med Inform ; 161: 104726, 2022 05.
Article in English | MEDLINE | ID: covidwho-1702743

ABSTRACT

BACKGROUND: The outbreak of the COVID-19 pandemic has led to the rapid adoption of novel telemedicine programs within the emergency department (ED) to minimize provider exposure and conserve personal protective equipment (PPE). In this study, we sought to assess how the adoption of telemedicine in the ED impacted clinical order patterns for patients with chest pain. We hypothesize that clinicians would rely more on imaging and laboratory workup for patients receiving telemedicine due to limitation in physical exams. METHODS: A single-center, retrospective, propensity score matched study was designed for patients presenting with chest pain at an ED. The study period was defined between April 1st, 2020 and September 30th, 2020. The frequency of the most frequent lab, imaging, and medication orders were compared. In addition, poisson regression analysis was performed to compare the overall number of orders between the two groups. RESULTS: 455 patients with chest pain who received telemedicine were matched to 455 similar patients without telemedicine with standardized mean difference < 0.1 for all matched covariates. The proportion of frequent lab, imaging, and medication orders were similar between the two groups. However, telemedicine patients received more orders overall (RR, 1.19, 95% CI, 1.11, 1.28, p-value < 0.001) as well as more imaging, lab, and nursing orders. The number of medication orders between the two groups remained similar. CONCLUSIONS: Frequent labs, imaging, and medications were ordered in similar proportions between the two cohorts. However, telemedicine patients had more orders placed overall. This study is an important objective assessment of the impact that telemedicine has upon clinical practice patterns and can guide future telemedicine implementation after the COVID-19 pandemic.


Subject(s)
COVID-19 , Telemedicine , Chest Pain/diagnosis , Chest Pain/epidemiology , Chest Pain/therapy , Emergency Service, Hospital , Humans , Pandemics , Practice Patterns, Physicians' , Retrospective Studies , Telemedicine/methods
6.
Transbound Emerg Dis ; 69(4): 2275-2286, 2022 Jul.
Article in English | MEDLINE | ID: covidwho-1304137

ABSTRACT

Angiotensin converting enzyme 2 (ACE2) is a host cell membrane protein (receptor) that mediates the binding of coronavirus, most notably SARS coronaviruses in the respiratory and gastrointestinal tracts. Although SARS-CoV-2 infection is mainly confined to humans, there have been numerous incidents of spillback (reverse zoonoses) to domestic and captive animals. An absence of information on the spatial distribution of ACE2 in animal tissues limits our understanding of host species susceptibility. Here, we describe the distribution of ACE2 using immunohistochemistry (IHC) on histological sections derived from carnivores, ungulates, primates and chiroptera. Comparison of mink (Neovison vison) and ferret (Mustela putorius furo) respiratory tracts showed substantial differences, demonstrating that ACE2 is present in the lower respiratory tract of mink but not ferrets. The presence of ACE2 in the respiratory tract in some species was much more restricted as indicated by limited immunolabelling in the nasal turbinate, trachea and lungs of cats (Felis catus) and only the nasal turbinate in the golden Syrian hamster (Mesocricetus auratus). In the lungs of other species, ACE2 could be detected on the bronchiolar epithelium of the sheep (Ovis aries), cattle (Bos taurus), European badger (Meles meles), cheetah (Acinonyx jubatus), tiger and lion (Panthera spp.). In addition, ACE2 was present in the nasal mucosa epithelium of the serotine bat (Eptesicus serotinus) but not in pig (Sus scrofa domestica), cattle or sheep. In the intestine, ACE2 immunolabelling was seen on the microvillus of enterocytes (surface of intestine) across various taxa. These results provide anatomical evidence of ACE2 expression in a number of species which will enable further understanding of host susceptibility and tissue tropism of ACE2 receptor-mediated viral infection.


Subject(s)
Angiotensin-Converting Enzyme 2 , COVID-19 , Receptors, Virus , Angiotensin-Converting Enzyme 2/metabolism , Animals , Animals, Wild , COVID-19/veterinary , Cat Diseases , Cats , Cattle , Cattle Diseases , Chiroptera , Ferrets , Livestock , Mink , Pets , Receptors, Virus/metabolism , SARS-CoV-2 , Sheep , Sheep Diseases , Spike Glycoprotein, Coronavirus/metabolism , Sus scrofa
7.
J Med Internet Res ; 23(7): e29240, 2021 07 26.
Article in English | MEDLINE | ID: covidwho-1302085

ABSTRACT

BACKGROUND: Telemedicine has been deployed by health care systems in response to the COVID-19 pandemic to enable health care workers to provide remote care for both outpatients and inpatients. Although it is reasonable to suspect telemedicine visits limit unnecessary personal contact and thus decrease the risk of infection transmission, the impact of the use of such technology on clinician workflows in the emergency department is unknown. OBJECTIVE: This study aimed to use a real-time locating system (RTLS) to evaluate the impact of a new telemedicine platform, which permitted clinicians located outside patient rooms to interact with patients who were under isolation precautions in the emergency department, on in-person interaction between health care workers and patients. METHODS: A pre-post analysis was conducted using a badge-based RTLS platform to collect movement data including entrances and duration of stay within patient rooms of the emergency department for nursing and physician staff. Movement data was captured between March 2, 2020, the date of the first patient screened for COVID-19 in the emergency department, and April 20, 2020. A new telemedicine platform was deployed on March 29, 2020. The number of entrances and duration of in-person interactions per patient encounter, adjusted for patient length of stay, were obtained for pre- and postimplementation phases and compared with t tests to determine statistical significance. RESULTS: There were 15,741 RTLS events linked to 2662 encounters for patients screened for COVID-19. There was no significant change in the number of in-person interactions between the pre- and postimplementation phases for both nurses (5.7 vs 7.0 entrances per patient, P=.07) and physicians (1.3 vs 1.5 entrances per patient, P=.12). Total duration of in-person interactions did not change (56.4 vs 55.2 minutes per patient, P=.74) despite significant increases in telemedicine videoconference frequency (0.6 vs 1.3 videoconferences per patient, P<.001 for change in daily average) and duration (4.3 vs 12.3 minutes per patient, P<.001 for change in daily average). CONCLUSIONS: Telemedicine was rapidly adopted with the intent of minimizing pathogen exposure to health care workers during the COVID-19 pandemic, yet RTLS movement data did not reveal significant changes for in-person interactions between staff and patients under investigation for COVID-19 infection. Additional research is needed to better understand how telemedicine technology may be better incorporated into emergency departments to improve workflows for frontline health care clinicians.


Subject(s)
COVID-19/diagnosis , COVID-19/prevention & control , Emergency Service, Hospital/organization & administration , Health Personnel/organization & administration , Telemedicine , Workflow , COVID-19/epidemiology , Cross Infection/prevention & control , Humans , Pandemics , SARS-CoV-2 , Time Factors
8.
West J Emerg Med ; 22(3): 552-560, 2021 Apr 28.
Article in English | MEDLINE | ID: covidwho-1266882

ABSTRACT

INTRODUCTION: In March 2020, shelter-in-place orders were enacted to attenuate the spread of coronavirus 2019 (COVID-19). Emergency departments (EDs) experienced unexpected and dramatic decreases in patient volume, raising concerns about exacerbating health disparities. METHODS: We queried our electronic health record to describe the overall change in visits to a two-ED healthcare system in Northern California from March-June 2020 compared to 2019. We compared weekly absolute numbers and proportional change in visits focusing on race/ethnicity, insurance, household income, and acuity. We calculated the z-score to identify whether there was a statistically significant difference in proportions between 2020 and 2019. RESULTS: Overall ED volume declined 28% during the study period. The nadir of volume was 52% of 2019 levels and occurred five weeks after a shelter-in-place order was enacted. Patient demographics also shifted. By week 4 (April 5), the proportion of Hispanic patients decreased by 3.3 percentage points (pp) (P = 0.0053) compared to a 6.2 pp increase in White patients (P = 0.000005). The proportion of patients with commercial insurance increased by 11.6 pp, while Medicaid visits decreased by 9.5 pp (P < 0.00001) at the initiation of shelter-in-place orders. For patients from neighborhoods <300% federal poverty levels (FPL), visits were -3.8 pp (P = 0.000046) of baseline compared to +2.9 pp (P = 0.0044) for patients from ZIP codes at >400% FPL the week of the shelter-in-place order. Overall, 2020 evidenced a consistently elevated proportion of high-acuity Emergency Severity Index (ESI) level 1 patients compared to 2019. Increased acuity was also demonstrated by an increase in the admission rate, with a 10.8 pp increase from 2019. Although there was an increased proportion of high-acuity patients, the overall census was decreased. CONCLUSION: Our results demonstrate changing ED utilization patterns circa the shelter-in-place orders. Those from historically vulnerable populations such as Hispanics, those from lower socioeconomic areas, and Medicaid users presented at disproportionately lower rates and numbers than other groups. As the pandemic continues, hospitals should use operations data to monitor utilization patterns by demographic, in addition to clinical indicators. Messaging about availability of emergency care and other services should include vulnerable populations to avoid exacerbating healthcare disparities.


Subject(s)
COVID-19/ethnology , Emergency Service, Hospital/statistics & numerical data , Healthcare Disparities/statistics & numerical data , Adolescent , Adult , Aged , Child , Child, Preschool , Economic Status/statistics & numerical data , Humans , Infant , Infant, Newborn , Insurance, Health/statistics & numerical data , Male , Medicaid/statistics & numerical data , Middle Aged , Pandemics , Retrospective Studies , SARS-CoV-2 , Severity of Illness Index , United States/epidemiology , Young Adult
9.
J Med Cases ; 11(11): 362-365, 2020 Nov.
Article in English | MEDLINE | ID: covidwho-1227210

ABSTRACT

Since the initial reported outbreak of coronavirus disease 2019 (COVID-19), many unique case reports have been published in the medical literature. Here we report a complicated clinical course of a young patient with COVID-19 who presented initially with recurrent autoimmune hemolytic anemia (AIHA). He subsequently developed bilateral pulmonary emboli, and ultimately succumbed to encephalitis and cryptococcemia in the context of being treated with high dose immunosuppression for the AIHA. Combining immunosuppression with active COVID-19 infection presents some truly challenging diagnostic and management scenarios which this case summarizes and highlights very well. Based on this case, we propose some strategies on how to approach these difficult decisions while also recognizing the significant gaps that exist in such an evolving topic. Lastly, this case also represents a potentially novel presentation of secondary fungal infection of the central nervous system (CNS) related to COVID-19.

10.
Viruses ; 13(4)2021 04 20.
Article in English | MEDLINE | ID: covidwho-1194712

ABSTRACT

SARS-CoV-2 virus was first detected in late 2019 and circulated globally, causing COVID-19, which is characterised by sub-clinical to severe disease in humans. Here, we investigate the serological antibody responses to SARS-CoV-2 infection during acute and convalescent infection using a cohort of (i) COVID-19 patients admitted to hospital, (ii) healthy individuals who had experienced 'COVID-19 like-illness', and (iii) a cohort of healthy individuals prior to the emergence of SARS-CoV-2. We compare SARS-CoV-2 specific antibody detection rates from four different serological methods, virus neutralisation test (VNT), ID Screen® SARS-CoV-2-N IgG ELISA, Whole Antigen ELISA, and lentivirus-based SARS-CoV-2 pseudotype virus neutralisation tests (pVNT). All methods were able to detect prior infection with COVID-19, albeit with different relative sensitivities. The VNT and SARS-CoV-2-N ELISA methods showed a strong correlation yet provided increased detection rates when used in combination. A pVNT correlated strongly with SARS-CoV-2 VNT and was able to effectively discriminate SARS-CoV-2 antibody positive and negative serum with the same efficiency as the VNT. Moreover, the pVNT was performed with the same level of discrimination across multiple separate institutions. Therefore, the pVNT is a sensitive, specific, and reproducible lower biosafety level alternative to VNT for detecting SARS-CoV-2 antibodies for diagnostic and research applications. Our data illustrate the potential utility of applying VNT or pVNT and ELISA antibody tests in parallel to enhance the sensitivity of exposure to infection.


Subject(s)
Antibodies, Viral/blood , COVID-19 Serological Testing/methods , COVID-19/diagnosis , SARS-CoV-2/immunology , Aged , Antibodies, Neutralizing/blood , COVID-19/blood , COVID-19/immunology , Cross Reactions , Enzyme-Linked Immunosorbent Assay , Female , Humans , Lentivirus/genetics , Male , Middle Aged , Neutralization Tests , Reproducibility of Results , SARS-CoV-2/isolation & purification , Sensitivity and Specificity , Spike Glycoprotein, Coronavirus/genetics , Spike Glycoprotein, Coronavirus/immunology
11.
J Virol ; 95(4)2021 01 28.
Article in English | MEDLINE | ID: covidwho-1075935

ABSTRACT

Swine influenza A virus (swIAV) infection causes substantial economic loss and disease burden in humans and animals. The 2009 pandemic H1N1 (pH1N1) influenza A virus is now endemic in both populations. In this study, we evaluated the efficacy of different vaccines in reducing nasal shedding in pigs following pH1N1 virus challenge. We also assessed transmission from immunized and challenged pigs to naive, directly in-contact pigs. Pigs were immunized with either adjuvanted, whole inactivated virus (WIV) vaccines or virus-vectored (ChAdOx1 and MVA) vaccines expressing either the homologous or heterologous influenza A virus hemagglutinin (HA) glycoprotein, as well as an influenza virus pseudotype (S-FLU) vaccine expressing heterologous HA. Only two vaccines containing homologous HA, which also induced high hemagglutination inhibitory antibody titers, significantly reduced virus shedding in challenged animals. Nevertheless, virus transmission from challenged to naive, in-contact animals occurred in all groups, although it was delayed in groups of vaccinated animals with reduced virus shedding.IMPORTANCE This study was designed to determine whether vaccination of pigs with conventional WIV or virus-vectored vaccines reduces pH1N1 swine influenza A virus shedding following challenge and can prevent transmission to naive in-contact animals. Even when viral shedding was significantly reduced following challenge, infection was transmissible to susceptible cohoused recipients. This knowledge is important to inform disease surveillance and control strategies and to determine the vaccine coverage required in a population, thereby defining disease moderation or herd protection. WIV or virus-vectored vaccines homologous to the challenge strain significantly reduced virus shedding from directly infected pigs, but vaccination did not completely prevent transmission to cohoused naive pigs.


Subject(s)
Influenza A Virus, H1N1 Subtype , Influenza Vaccines/administration & dosage , Orthomyxoviridae Infections/transmission , Swine Diseases/transmission , Virus Shedding , Adjuvants, Immunologic/administration & dosage , Animals , Female , Influenza A Virus, H1N1 Subtype/immunology , Influenza A Virus, H1N1 Subtype/isolation & purification , Orthomyxoviridae Infections/prevention & control , Swine , Swine Diseases/prevention & control , Vaccination , Vaccines, Attenuated/administration & dosage , Vaccines, Inactivated/administration & dosage
12.
Viruses ; 13(1)2021 Jan 15.
Article in English | MEDLINE | ID: covidwho-1067778

ABSTRACT

Ferrets were experimentally inoculated with SARS-CoV-2 (severe acute respiratory syndrome (SARS)-related coronavirus 2) to assess infection dynamics and host response. During the resulting subclinical infection, viral RNA was monitored between 2 and 21 days post-inoculation (dpi), and reached a peak in the upper respiratory cavity between 4 and 6 dpi. Viral genomic sequence analysis in samples from three animals identified the Y453F nucleotide substitution relative to the inoculum. Viral RNA was also detected in environmental samples, specifically in swabs of ferret fur. Microscopy analysis revealed viral protein and RNA in upper respiratory tract tissues, notably in cells of the respiratory and olfactory mucosae of the nasal turbinates, including olfactory neuronal cells. Antibody responses to the spike and nucleoprotein were detected from 21 dpi, but virus-neutralizing activity was low. A second intranasal inoculation (re-exposure) of two ferrets after a 17-day interval did not produce re-initiation of viral RNA shedding, but did amplify the humoral response in one animal. Therefore, ferrets can be experimentally infected with SARS-CoV-2 to model human asymptomatic infection.


Subject(s)
Asymptomatic Diseases , COVID-19/virology , Disease Models, Animal , SARS-CoV-2/physiology , Animals , Antibodies, Viral/blood , Antibodies, Viral/immunology , COVID-19/diagnosis , COVID-19/pathology , COVID-19/transmission , Female , Ferrets , Genome, Viral/genetics , Mutation , Nasal Mucosa/virology , RNA, Viral/genetics , SARS-CoV-2/isolation & purification , Viral Load , Virus Shedding
13.
Viruses ; 13(2)2021 02 03.
Article in English | MEDLINE | ID: covidwho-1060766

ABSTRACT

The long-term control strategy of SARS-CoV-2 and other major respiratory viruses needs to include antivirals to treat acute infections, in addition to the judicious use of effective vaccines. Whilst COVID-19 vaccines are being rolled out for mass vaccination, the modest number of antivirals in use or development for any disease bears testament to the challenges of antiviral development. We recently showed that non-cytotoxic levels of thapsigargin (TG), an inhibitor of the sarcoplasmic/endoplasmic reticulum (ER) Ca2+ ATPase pump, induces a potent host innate immune antiviral response that blocks influenza A virus replication. Here we show that TG is also highly effective in blocking the replication of respiratory syncytial virus (RSV), common cold coronavirus OC43, SARS-CoV-2 and influenza A virus in immortalized or primary human cells. TG's antiviral performance was significantly better than remdesivir and ribavirin in their respective inhibition of OC43 and RSV. Notably, TG was just as inhibitory to coronaviruses (OC43 and SARS-CoV-2) and influenza viruses (USSR H1N1 and pdm 2009 H1N1) in separate infections as in co-infections. Post-infection oral gavage of acid-stable TG protected mice against a lethal influenza virus challenge. Together with its ability to inhibit the different viruses before or during active infection, and with an antiviral duration of at least 48 h post-TG exposure, we propose that TG (or its derivatives) is a promising broad-spectrum inhibitor against SARS-CoV-2, OC43, RSV and influenza virus.


Subject(s)
Antiviral Agents/pharmacology , Betacoronavirus/drug effects , Coronavirus OC43, Human/drug effects , Influenza A Virus, H1N1 Subtype/drug effects , Respiratory Syncytial Virus, Human/drug effects , SARS-CoV-2/drug effects , Thapsigargin/pharmacology , Animals , Antiviral Agents/therapeutic use , Betacoronavirus/physiology , Cell Line , Cell Line, Tumor , Cells, Cultured , Coronavirus OC43, Human/physiology , Endoplasmic Reticulum Stress , Humans , Influenza A Virus, H1N1 Subtype/physiology , Mice , Microbial Sensitivity Tests , Orthomyxoviridae Infections/drug therapy , Orthomyxoviridae Infections/virology , Respiratory Syncytial Virus, Human/physiology , Ribavirin/pharmacology , SARS-CoV-2/physiology , Thapsigargin/therapeutic use , Virus Replication/drug effects
14.
J Virol Methods ; 289: 114048, 2021 03.
Article in English | MEDLINE | ID: covidwho-988686

ABSTRACT

We describe the optimisation of a simplified sample preparation method which permits rapid and direct detection of SARS-CoV-2 RNA within saliva, using reverse-transcription loop-mediated isothermal amplification (RT-LAMP). Treatment of saliva samples prior to RT-LAMP by dilution 1:1 in Mucolyse™, followed by dilution in 10 % (w/v) Chelex© 100 Resin and a 98 °C heat step for 2 min enabled detection of SARS-CoV-2 RNA in positive saliva samples. Using RT-LAMP, SARS-CoV-2 RNA was detected in as little as 05:43 min, with no amplification detected in 3097 real-time reverse transcription PCR (rRT-PCR) negative saliva samples from staff tested within a service evaluation study, or for other respiratory pathogens tested (n = 22). Saliva samples can be collected non-invasively, without the need for skilled staff and can be obtained from both healthcare and home settings. Critically, this approach overcomes the requirement for, and validation of, different swabs and the global bottleneck in obtaining access to extraction robots and reagents to enable molecular testing by rRT-PCR. Such testing opens the possibility of public health approaches for effective intervention during the COVID-19 pandemic through regular SARS-CoV-2 testing at a population scale, combined with isolation and contact tracing.


Subject(s)
COVID-19 Testing/methods , COVID-19/diagnosis , Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , SARS-CoV-2/isolation & purification , Saliva/virology , Specimen Handling/methods , Humans , RNA, Viral/analysis
15.
Sci Rep ; 10(1): 21894, 2020 12 14.
Article in English | MEDLINE | ID: covidwho-977275

ABSTRACT

The rapid emergence of SARS-CoV-2, the causative agent of COVID-19, and its dissemination globally has caused an unprecedented strain on public health. Animal models are urgently being developed for SARS-CoV-2 to aid rational design of vaccines and therapeutics. Immunohistochemistry and in situ hybridisation techniques that facilitate reliable and reproducible detection of SARS-CoV and SARS-CoV-2 viral products in formalin-fixed paraffin-embedded (FFPE) specimens would be of great utility. A selection of commercial antibodies generated against SARS-CoV spike protein and nucleoprotein, double stranded RNA, and RNA probe for spike genes were evaluated for the ability to detect FFPE infected cells. We also tested both heat- and enzymatic-mediated virus antigen retrieval methods to determine the optimal virus antigen recovery as well as identifying alternative retrieval methods to enable flexibility of IHC methods. In addition to using native virus infected cells as positive control material, the evaluation of non-infected cells expressing coronavirus (SARS, MERS) spike as a biosecure alternative to assays involving live virus was undertaken. Optimized protocols were successfully applied to experimental animal-derived tissues. The diverse techniques for virus detection and control material generation demonstrated in this study can be applied to investigations of coronavirus pathogenesis and therapeutic research in animal models.


Subject(s)
Antigens, Viral/immunology , COVID-19 Testing , COVID-19 , Immunohistochemistry , SARS-CoV-2/isolation & purification , Animals , Antibodies, Monoclonal/immunology , Antibodies, Viral/immunology , COVID-19/diagnosis , COVID-19/virology , Chlorocebus aethiops , Ferrets , In Situ Hybridization , RNA Probes/immunology , Severe acute respiratory syndrome-related coronavirus/isolation & purification , Vero Cells
16.
Viruses ; 12(12)2020 12 09.
Article in English | MEDLINE | ID: covidwho-965146

ABSTRACT

Emerging infectious diseases are of great concern to public health, as highlighted by the ongoing coronavirus disease 2019 (COVID-19) pandemic. Such diseases are of particular danger during mass gathering and mass influx events, as large crowds of people in close proximity to each other creates optimal opportunities for disease transmission. The Hashemite Kingdom of Jordan and the Kingdom of Saudi Arabia are two countries that have witnessed mass gatherings due to the arrival of Syrian refugees and the annual Hajj season. The mass migration of people not only brings exotic diseases to these regions but also brings new diseases back to their own countries, e.g., the outbreak of MERS in South Korea. Many emerging pathogens originate in bats, and more than 30 bat species have been identified in these two countries. Some of those bat species are known to carry viruses that cause deadly diseases in other parts of the world, such as the rabies virus and coronaviruses. However, little is known about bats and the pathogens they carry in Jordan and Saudi Arabia. Here, the importance of enhanced surveillance of bat-borne infections in Jordan and Saudi Arabia is emphasized, promoting the awareness of bat-borne diseases among the general public and building up infrastructure and capability to fill the gaps in public health preparedness to prevent future pandemics.


Subject(s)
Chiroptera/virology , Communicable Diseases, Emerging/epidemiology , Coronavirus/isolation & purification , Public Health , Zoonoses/epidemiology , Animals , COVID-19 , Communicable Diseases, Emerging/virology , Coronavirus/classification , Coronavirus/pathogenicity , Disease Outbreaks/prevention & control , Humans , Jordan , Saudi Arabia , Zoonoses/transmission , Zoonoses/virology
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