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2.
Life Sci Alliance ; 6(1)2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36241425

RESUMO

New therapeutic targets are a valuable resource for treatment of SARS-CoV-2 viral infection. Genome-wide association studies have identified risk loci associated with COVID-19, but many loci are associated with comorbidities and are not specific to host-virus interactions. Here, we identify and experimentally validate a link between reduced expression of EXOSC2 and reduced SARS-CoV-2 replication. EXOSC2 was one of the 332 host proteins examined, all of which interact directly with SARS-CoV-2 proteins. Aggregating COVID-19 genome-wide association studies statistics for gene-specific eQTLs revealed an association between increased expression of EXOSC2 and higher risk of clinical COVID-19. EXOSC2 interacts with Nsp8 which forms part of the viral RNA polymerase. EXOSC2 is a component of the RNA exosome, and here, LC-MS/MS analysis of protein pulldowns demonstrated interaction between the SARS-CoV-2 RNA polymerase and most of the human RNA exosome components. CRISPR/Cas9 introduction of nonsense mutations within EXOSC2 in Calu-3 cells reduced EXOSC2 protein expression and impeded SARS-CoV-2 replication without impacting cellular viability. Targeted depletion of EXOSC2 may be a safe and effective strategy to protect against clinical COVID-19.


Assuntos
COVID-19 , SARS-CoV-2 , COVID-19/genética , Cromatografia Líquida , Códon sem Sentido , RNA Polimerases Dirigidas por DNA/genética , Complexo Multienzimático de Ribonucleases do Exossomo/genética , Estudo de Associação Genômica Ampla , Humanos , RNA Viral/metabolismo , Proteínas de Ligação a RNA/genética , SARS-CoV-2/genética , Espectrometria de Massas em Tandem , Proteínas do Complexo da Replicase Viral , Replicação Viral/genética
3.
Adv Neurobiol ; 29: 449-477, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36255684

RESUMO

A number of viruses that have caused wide spread concern e.g. Ebola, Zika, and SARS-CoV2 (severe acute respiratory syndrome coronavirus 2 also known as COVID 19) have at various times, become newsworthy as a result of being newly discovered, mutations enabling them to more efficiently infect humans or modern modes of transportation moving them to areas with naive, susceptible populations. As more is learned about the mechanisms whereby these pathogens enter human cells it has become increasingly evident that carbohydrates expressed on the surface of either target cells or the pathogens themselves are essential. Variability in carbohydrate structures as well as the presence of carbohydrate binding receptors (lectins) provides a plethora of potential binding interactions by which infection of cells can occur. Identification of specific lipid- or protein-associated carbohydrates essential for infection provides support for research being done to develop carbohydrate related inhibitors of those interactions. This chapter (1) discusses scenarios for how carbohydrates affect the ability of specific infectious agents to interact with neural cells, (2) gives examples of problems that may result from development of antibodies to carbohydrate antigens found on pathogens that are similar to epitopes expressed on mammalian cells, and (3) provides examples of approaches either in use or under consideration for translational uses of this information.


Assuntos
COVID-19 , Infecção por Zika virus , Zika virus , Animais , Humanos , SARS-CoV-2 , RNA Viral , Carboidratos/química , Sítios de Ligação , Epitopos , Lectinas , Lipídeos , Infecção por Zika virus/tratamento farmacológico , Mamíferos
4.
Methods Mol Biol ; 2570: 205-222, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36156785

RESUMO

RNA molecules are essential for carrying genetic information and regulating gene expression in most organisms including human pathogenic RNA and relate retro viruses. Targeting viral RNA (vRNA) structures provide broad opportunities to develop chemical tools to probe molecular virology and to discover novel targets for therapeutic intervention. An increasing number of RNA binding small molecules are being identified, stimulating increased interests in small molecule drug discovery for RNA targets. In this chapter, we describe protocols to characterize and robustly validate vRNA-small molecule (vRNA-sm) interactions starting from vRNA sample preparation, followed by small molecule screening against vRNA targets and finally to validating the vRNA-sm interactions via NMR spectroscopy and calorimetric titrations.


Assuntos
RNA Viral , Biofísica , Calorimetria , Humanos , RNA Viral/química
5.
J Hazard Mater ; 441: 129848, 2023 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-36067562

RESUMO

Wastewater-based epidemiology (WBE) has been considered as a promising approach for population-wide surveillance of coronavirus disease 2019 (COVID-19). Many studies have successfully quantified severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA concentration in wastewater (CRNA). However, the correlation between the CRNA and the COVID-19 clinically confirmed cases in the corresponding wastewater catchments varies and the impacts of environmental and other factors remain unclear. A systematic review and meta-analysis were conducted to identify the correlation between CRNA and various types of clinically confirmed case numbers, including prevalence and incidence rates. The impacts of environmental factors, WBE sampling design, and epidemiological conditions on the correlation were assessed for the same datasets. The systematic review identified 133 correlation coefficients, ranging from -0.38 to 0.99. The correlation between CRNA and new cases (either daily new, weekly new, or future cases) was stronger than that of active cases and cumulative cases. These correlation coefficients were potentially affected by environmental and epidemiological conditions and WBE sampling design. Larger variations of air temperature and clinical testing coverage, and the increase of catchment size showed strong negative impacts on the correlation between CRNA and COVID-19 case numbers. Interestingly, the sampling technique had negligible impact although increasing the sampling frequency improved the correlation. These findings highlight the importance of viral shedding dynamics, in-sewer decay, WBE sampling design and clinical testing on the accurate back-estimation of COVID-19 case numbers through the WBE approach.


Assuntos
COVID-19 , COVID-19/epidemiologia , Humanos , RNA Viral/genética , SARS-CoV-2/genética , Águas Residuárias , Vigilância Epidemiológica Baseada em Águas Residuárias
6.
Methods Mol Biol ; 2578: 199-208, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36152289

RESUMO

Flavivirus are the most alarming prevalent viruses worldwide due to its vast impact on public health. Most early symptoms of diseases caused by Flavivirus are similar among each other and to other febrile illnesses making the clinical differential diagnosis challenging. In addition, due to cross-reactivity and a relatively limited persistence of viral RNA in infected individuals, the current available diagnosis strategies fail to efficiently provide a differential viral identification. In this context, virus-specific tests are essential to improve patient care, as well as to facilitate disease surveillance and the effective control of transmission. Here, we describe the use of protein microarrays as an effective tool for screening peptides differentially recognized by anti-Yellow Fever virus antibodies induced by vaccination or by natural viral infection.


Assuntos
Flavivirus , Anticorpos Antivirais , Reações Cruzadas , Flavivirus/genética , Humanos , Peptídeos , RNA Viral/genética
7.
J Environ Sci (China) ; 125: 843-850, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36375966

RESUMO

With a unique and large size of testing results of 1,842 samples collected from 12 wastewater treatment plants (WWTP) for 14 months through from low to high prevalence of COVID-19, the sensitivity of RT-qPCR detection of SARS-CoV-2 RNA in wastewater that correspond to the communities was computed by using Probit analysis. This study determined the number of new COVID-19 cases per 100,000 population required to detect SARS-CoV-2 RNA in wastewater at defined probabilities and provided an evidence-based framework of wastewater-based epidemiology surveillance (WBE). Input data were positive and negative test results of SARS-CoV-2 RNA in wastewater samples and the corresponding new COVID-19 case rates per 100,000 population served by each WWTP. The analyses determined that RT-qPCR-based SARS-CoV-2 RNA detection threshold at 50%, 80% and 99% probability required a median of 8 (range: 4-19), 18 (9-43), and 38 (17-97) of new COVID-19 cases /100,000, respectively. Namely, the positive detection rate at 50%, 80% and 99% probability were 0.01%, 0.02%, and 0.04% averagely for new cases in the population. This study improves understanding of the performance of WBE SARS-CoV-2 RNA detection using the large datasets and prolonged study period. Estimated COVID-19 burden at a community level that would result in a positive detection of SARS-CoV-2 in wastewater is critical to support WBE application as a supplementary warning/monitoring system for COVID-19 prevention and control.


Assuntos
COVID-19 , Humanos , COVID-19/epidemiologia , SARS-CoV-2/genética , Águas Residuárias/análise , RNA Viral/genética , RNA Viral/análise , Alberta/epidemiologia
9.
Sci Total Environ ; 856(Pt 1): 158966, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36162583

RESUMO

During the coronavirus disease 2019 (COVID-19) pandemic, wastewater-based epidemiology (WBE) attracted attention as an objective and comprehensive indicator of community infection that does not require individual inspection. Although several severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection methods from wastewater have been developed, there are obstacles to their social implementation. In this study, we developed the COPMAN (Coagulation and Proteolysis method using Magnetic beads for detection of Nucleic acids in wastewater), an automatable method that can concentrate and detect multiple types of viruses from a limited volume (∼10 mL) of wastewater. The COPMAN consists of a high basicity polyaluminum chloride (PAC) coagulation process, magnetic bead-based RNA purification, and RT-preamplification, followed by qPCR. A series of enzymes exhibiting a high tolerance to PCR inhibitors derived from wastewater was identified and employed in the molecular detection steps in the COPMAN. We compared the detectability of viral RNA from 10-mL samples of virus-spiked (heat-inactivated SARS-CoV-2 and intact RSV) or unspiked wastewater by the COPMAN and other methods (PEG-qPCR, UF-qPCR, and EPISENS-S). The COPMAN was the most efficient for detecting spiked viruses from wastewater, detecting the highest level of pepper mild mottle virus (PMMoV), a typical intrinsic virus in human stool, from wastewater samples. The COPMAN also successfully detected indigenous SARS-CoV-2 RNA from 12 samples of wastewater at concentrations of 2.2 × 104 to 5.4 × 105 copies/L, during initial stages of an infection wave in the right and the left bank of the Sagami River in Japan (0.65 to 11.45 daily reported cases per 100,000 people). These results indicate that the COPMAN is suitable for detection of multiple pathogens from small volume of wastewater in automated stations.


Assuntos
COVID-19 , Ácidos Nucleicos , Vírus , Humanos , SARS-CoV-2/genética , RNA Viral , Águas Residuárias , COVID-19/diagnóstico
10.
Environ Res ; 216(Pt 1): 114089, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36007572

RESUMO

Several studies have proposed that environmental factors influencing human wellbeing, such as chronic exposures to high levels of particulate matter, could indirectly or even directly affect also the severity of COVID-19 disease in case of infection by novel coronavirus SARS-COV2. This study has investigated the association between COVID-19 infections, hospitalizations or deaths and the extension of public green areas (km2 per 100,000 based on OECD data of 2014), an indicator that has been chosen as independent endpoint variable to test the research hypothesis in 10 Italian and 8 Spanish Provinces with more than 500.000 inhabitants, including capitals (Rome and Madrid) and bigger cities (Bologna, Catania, Florence, Genoa, Milan, Naples, Palermo, Turin and Venice for Italy; Barcelona, Valencia, Seville, Zaragoza, Malaga, Las Palmas and Bilbao for Spain). Two different methodologies have been applied: a bottom-up approach was applied to Spanish institutional data concerning contagions/hospitalizations/deaths and the extent of public green areas for each responder to an official questionnaire in the frame of a nationwide survey (with detailed data granularity per province) containing specific georeferenced information; a top-down approach was used for Italy, starting from the official figures of contagions/hospitalizations/deaths of each province and linking them to the OECD statistics about the extension of public green areas in the different areas. Linear and generalized models were used for statistical analyses including also PM2.5 in a multivariate approach (with annual average concentrations from official air quality monitoring stations) and were able to adjust for the different number inhabitants living in each province, in order to take into account the difference in contagion dynamics related to the different density of population. The results obtained for Spain are consistent with those observed for Italy, as for both countries, it has clearly emerged a statistically significant association between COVID-19 clinical features (contagions, hospitalizations, and deaths) and the extension of public green areas, as well as the annual average concentrations of PM2.5 (with this latter variable loosing statistical significance in some province). Therefore, the extension of public green areas and air pollution seem to have a high correlation with COVID-19 severity.


Assuntos
Poluentes Atmosféricos , Poluição do Ar , COVID-19 , Humanos , COVID-19/epidemiologia , Espanha/epidemiologia , SARS-CoV-2 , Estudos Retrospectivos , Poluentes Atmosféricos/análise , RNA Viral , Poluição do Ar/análise , Material Particulado/análise , Itália/epidemiologia
11.
Sci Total Environ ; 857(Pt 2): 159401, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36240930

RESUMO

Understanding the persistence of SARS-CoV-2 biomarkers in wastewater should guide wastewater-based epidemiology users in selecting best RNA biomarkers for reliable detection of the virus during current and future waves of the pandemic. In the present study, the persistence of endogenous SARS-CoV-2 were assessed during one month for six different RNA biomarkers and for the pepper mild mottle virus (PMMoV) at three different temperatures (4, 12 and 20 °C) in one wastewater sample. All SARS-CoV-2 RNA biomarkers were consistently detected during 6 days at 4° and differences in signal persistence among RNA biomarkers were mostly observed at 20 °C with N biomarkers being globally more persistent than RdRP, E and ORF1ab ones. SARS-CoV-2 signal persistence further decreased in a temperature dependent manner. At 12 and 20 °C, RNA biomarker losses of 1-log10 occurred on average after 6 and 4 days, and led to a complete signal loss after 13 and 6 days, respectively. Besides the effect of temperature, SARS-CoV-2 RNA signals were more persistent in the particulate phase compared to the aqueous one. Finally, PMMoV RNA signal was highly persistent in both phases and significantly differed from that of SARS-CoV-2 biomarkers. We further provide a detailed overview of the latest literature on SARS-CoV-2 and PMMoV decay rates in sewage matrices.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , Vigilância Epidemiológica Baseada em Águas Residuárias , Águas Residuárias , Temperatura , RNA Viral , COVID-19/epidemiologia
12.
Sci Total Environ ; 857(Pt 2): 159358, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36240928

RESUMO

Wastewater-based epidemiology provides a conceptual framework for the evaluation of the prevalence of public health related biomarkers. In the context of the Coronavirus disease-2019, wastewater monitoring emerged as a complementary tool for epidemic management. In this study, we evaluated data from six wastewater treatment plants in the region of Saxony, Germany. The study period lasted from February to December 2021 and covered the third and fourth regional epidemic waves. We collected 1065 daily composite samples and analyzed SARS-CoV-2 RNA concentrations using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Regression models quantify the relation between RNA concentrations and disease prevalence. We demonstrated that the relation is site and time specific. Median loads per diagnosed case differed by a factor of 3-4 among sites during both waves and were on average 45 % higher during the third wave. In most cases, log-log-transformed data achieved better regression performance than non-transformed data and local calibration outperformed global models for all sites. The inclusion of lag/lead time, discharge and detection probability improved model performance in all cases significantly, but the importance of these components was also site and time specific. In all cases, models with lag/lead time and log-log-transformed data obtained satisfactory goodness-of-fit with adjusted coefficients of determination higher than 0.5. Back-estimation of testing efficiency from wastewater data confirmed state-wide prevalence estimation from individual testing statistics, but revealed pronounced differences throughout the epidemic waves and among the different sites.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , Águas Residuárias/análise , COVID-19/epidemiologia , RNA Viral , Prevalência , Biomarcadores
13.
Food Microbiol ; 109: 104114, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36309428

RESUMO

Hepatitis E virus (HEV) is the causative agent of hepatitis E. Some of the rise in hepatitis E infection in China may be linked to undercooked pork. In this study, we established a reverse transcription droplet digital PCR (RT-ddPCR) method to detect HEV in raw pork livers. The detection limit of the assay for HEV RNA was as low as 1.81 copies/µL. The suggested approach was validated on 14 samples, demonstrating greater sensitivity, specificity, and anti-interference performance features than RT-qPCR. Furthermore, we amplified the partial ORF2 gene by nested RT-PCR and sequenced for the HEV RNA positive samples. The prevalence of HEV in all collected samples was 2.24% (14/626), and the viral load was between 8.0 copies/µL and 8975 copies/µL. Specifically, the virus was detected in 10.62% (12/113) of the samples collected from the bio-safety disposal centers for dead livestock and poultry, in 0.67% (2/300) of the samples collected from the slaughterhouses, and none of the samples collected from the retail markets was HEV RNA positive. The subsequent phylogenetic analysis revealed that all HEV isolates belonged to the subtype 4d, which is one of the most common subtypes in northern China.


Assuntos
Vírus da Hepatite E , Hepatite E , Carne de Porco , Carne Vermelha , Doenças dos Suínos , Animais , Suínos , Humanos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Carne Vermelha/análise , Filogenia , RNA Viral/genética , RNA Viral/análise , Doenças dos Suínos/epidemiologia , Genótipo
14.
Ann Ig ; 35(1): 112-120, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36222607

RESUMO

Background: The COVID-19 emergency has highlighted the importance of prevention systems and environ-mental microbiological monitoring as fundamental elements in the response to epidemics and other such threats to individual and collective health. The use of automated "No-touch" room disinfection systems eliminates or reduces the dependence on operators, thus allowing an improvement in the effectiveness of terminal disinfection. Study design: In the present study, we focused on possible SARS-CoV-2 contamination of surfaces of com-mercial services, and the effectiveness of ozone treatment on the virus. Methods: Analyses were conducted on 4-7 October and 27-30 December 2021 in four supermarkets in an Apulian city; supermarkets A and B were equipped with an ozonisation system, while C and D were without any environmental remediation. Results: SARS-CoV-2 RNA was detected by real-time RT-PCR only in December, in 6% of the surfaces tested, and all examined samples were found to be negative after viral culture, since no cytopathic effect was observed. A statistically significant difference emerged from the comparison of October vs. December (p = 0.0289), but no statistically significant difference (p = 0.6777) emerged from the comparison between supermarkets with and without the ozonisation system. Conclusions: Although no important changes were observed by treating the environments with ozonisation systems, further studies are needed to validate the effectiveness of environmental treatments with airborne disinfectants.


Assuntos
COVID-19 , Ozônio , Humanos , SARS-CoV-2 , RNA Viral , Desinfecção , Ozônio/farmacologia
15.
Ann Ig ; 35(1): 75-83, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-35532052

RESUMO

Background: Vaccination has saved millions of lives through the protection of individuals and populations from communicable diseases. Vaccine hesitancy, defined as the delay in acceptance or refusal of vaccines despite the availability of vaccination services, has become a growing global concern. The objective of this study was to investigate parents'/caregivers' hesitancy toward childhood vaccination and its predictors in Albania. Study design: A cross-sectional survey was conducted. Methods: The data comes from a survey conducted on a sample of parents/caregivers (89.6% mothers) of children aged 6 months to 8 years at health care vaccination centers in seven Albanian cities from December 2020 to February 2021. Parents/caregivers (one per child) were interviewed by trained healthcare staff using a standardized questionnaire on six main content domains, including immunization behavior, beliefs about vaccine safety and efficacy, attitudes about vaccines, vaccination confidence, estimation of vaccine delay, and the intention to immunize children against SARS-CoV-2, and a self-reported hesitancy. The Albanian Ministry of Health approved the questionnaire, after it was translated, validated and adapted to the local setting. Statistical analyses included independent sample t-tests (p<0.05) and a logistic regression (OR; 95% C.I.). Results: A total of 475 parents/caregivers of children aged from 6 months to 8 years, attending childhood vaccination in public health services, were interviewed. To the question "how hesitant you are about childhood vaccination", a high number of parents/caregivers (46%) responded that they do not feel hesitant at all, and 32% were not hesitant, a small number of parents/caregivers said they are very hesitant (5%) or somewhat hesitant (12%). A binary logistic model was fitted to the data to test the hypothesis regarding the relationship between parental vaccine hesitancy and possible predictors. A lower parental attitude toward childhood vaccines (OR = 3.7; 95% C.I. 1.102-12.421), a health center with a high vaccine delay (OR = 2.878; C.I. 95% 1.735-4.773), and low confidence in health staff information (OR = 2.042; 95% C.I. 1.156-3.605) were all independent predictors of parental vaccine hesitancy. Regarding intention to vaccinate children against COVID-19, when available, nearly 75% of parents/caregivers showed hesitancy. Conclusions: Our results highlighted the role of positive parents'/caregivers' attitudes toward childhood vaccines followed by high staff confidence and good health center organization in order to deal with vaccine hesitancy, particularly for traditional and well-known childhood vaccines. Nevertheless, the hesitancy can be a critical barrier for childhood vaccination when we have to introduce a new vaccine, as is demonstrated in the recent vaccination campaign against the ongoing pandemic of SARS-CoV2.


Assuntos
COVID-19 , Vacinas , Criança , Feminino , Humanos , Cuidadores , Estudos Transversais , Hesitação Vacinal , Albânia , RNA Viral , Conhecimentos, Atitudes e Prática em Saúde , COVID-19/prevenção & controle , SARS-CoV-2 , Pais , Vacinação
16.
Talanta ; 252: 123835, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-35985194

RESUMO

In this paper we present a new method for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), targeting a specific region "N gene." Under isothermal reaction conditions, we integrated ligation (Lig; high selectivity) and recombinase polymerase amplification (RPA; high sensitivity) processes, obtaining a robust method of detection. For point-of-care testing, we incorporated our laboratory-produced pyrophosphate ion (PPi)-sensing probe (PK-probe) for colorimetric analysis of the reaction. The total detection system was efficient and effective at diagnosing this RNA virus-mediated disease rapidly (30 min). In a full-genome SARS-CoV-2 study, our PK-probe/Lig-RPA system functioned with a limit of detection of 1160 copies/ml, with a single-mismatch level of selectively, and it was highly selective even in the presence of bacterial genomes commonly found in the human mouth and nose. This robust, straightforward, selective, efficient, and ultrasensitive colorimetric detection method, with potential for point-of-care analysis, should also be effective in detecting a diverse range of other RNA-based diseases.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , Recombinases , Colorimetria , Técnicas de Amplificação de Ácido Nucleico/métodos , COVID-19/diagnóstico , Sensibilidade e Especificidade , RNA Viral
17.
Phytomedicine ; 108: 154496, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36288651

RESUMO

BACKGROUND: Jingyin granules (JY), one patented Chinese herbal formula, have been advised for treating coronavirus disease 2019 (COVID-19) in China. As of now, the safety and effectiveness of JY in treating COVID-19 patients were still to be evaluated. PURPOSE: To investigate the safety and clinical effectiveness of JY in treating mild COVID-19 patients. STUDY DESIGN: We carried out a prospective cohort study, as the highly infectious COVID-19 omicron variant ranged in Shanghai (ClinicalTrial.gov registration number: ChiCTR2200058692). METHODS: Participants infected with COVID-19, who were diagnosed as mild cases, were assigned to receive either JY (JY group) or traditional Chinese medicine placebo (placebo group) orally for 7 days. The primary clinical indicators were the RNA negative conversion rate (NCR) and the incidence of severe cases. The secondary clinical indicators were the negative conversion time (NCT), inpatient length of stay (ILOS), and the disappearance rates of clinical symptoms. RESULTS: Nine hundred participants were recruited in this clinical trial study, and 830 patients met the eligibility criteria. Seven hundred and ninety-one patients, accomplished the following-up assessment, including 423 cases of JY group and 368 cases of placebo group. NCR in JY group at 7-day posttreatment was considerably greater compared with placebo group (89.8% [380/423] vs 82.6% [304/368], P = 0.003). None of the patients with mild COVID-19 developed into severe cases. The median NCT of SARS-CoV-2 and ILOS in JY group were lesser than that in placebo group (4.0 [3.0,6.0]vs 5.0 [4.0,7.0] days, P < 0.001; 6.0 [4.0, 8.0] vs 7.0 [5.0, 9.0] days, P < 0.001). In both groups, the obvious improvement in clinical symptoms was observed, but the difference was not significant. In the subgroup of age ≤ 60 years, JY promoted SARS-CoV-2 RNA negative conversion (HR=1.242; 95% CI: 1.069-1.444, P < 0.001). No patients in both groups were reported as the case of serious adverse event. CONCLUSION: JY maybe the potential medicine for treating mild COVID-19 patients, which had beneficial effects on increasing NCR, and shortening NCT and ILOS.


Assuntos
COVID-19 , Humanos , Pessoa de Meia-Idade , China , COVID-19/tratamento farmacológico , Medicamentos sem Prescrição , Estudos Prospectivos , RNA Viral , SARS-CoV-2 , Resultado do Tratamento
18.
J Virol Methods ; 311: 114645, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36332716

RESUMO

Wastewater monitoring for severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), the virus responsible for the global coronavirus disease 2019 (COVID-19) pandemic, has highlighted the need for methodologies capable of assessing viral prevalence during periods of low population infection. To address this need, two volumetrically different, methodologically similar concentration approaches were compared for their abilities to detect viral nucleic acid and infectious SARS-CoV-2 signal from primary influent samples. For Method 1, 2 L of SARS-CoV-2 seeded wastewater was evaluated using a dead-end hollow fiber ultrafilter (D-HFUF) for primary concentration, followed by the CP Select™ for secondary concentration. For Method 2, 100 mL of SARS-CoV-2 seeded wastewater was evaluated using the CP Select™ procedure. Following D-HFUF concentration (Method 1), significantly lower levels of infectious SARS-CoV-2 were lost (P value range: 0.0398-0.0027) compared to viral gene copy (GC) levels detected by the US Centers for Disease Control (CDC) N1 and N2 reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) assays. Subsamples at different steps in the concentration process were also taken to better characterize the losses of SARS-CoV-2 during the concentration process. During the centrifugation step (prior to CP Select™ concentration), significantly higher losses (P value range: 0.0003 to <0.0001) occurred for SARS-CoV-2 GC levels compared to infectious virus for Method 1, while between the methods, significantly higher infectious viral losses were observed for Method 2 (P = 0.0002). When analyzing overall recovery of endogenous SARS-CoV-2 in wastewater samples, application of Method 1 improved assay sensitivities (P = <0.0001) compared with Method 2; this was especially evident during periods of lower COVID-19 case rates within the sewershed. This study describes a method which can successfully concentrate infectious SARS-CoV-2 and viral RNA from wastewater. Moreover, we demonstrated that large volume wastewater concentration provides additional sensitivity needed to improve SARS-CoV-2 detection, especially during low levels of community disease prevalence.


Assuntos
COVID-19 , Vírus , Humanos , SARS-CoV-2/genética , COVID-19/diagnóstico , Águas Residuárias , Pandemias , RNA Viral/genética
19.
Sci Total Environ ; 857(Pt 1): 159188, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36202365

RESUMO

Genomic footprints of pathogens shed by infected individuals can be traced in environmental samples, which can serve as a noninvasive method of infectious disease surveillance. The research evaluates the efficacy of environmental monitoring of SARS-CoV-2 RNA in air, surface swabs and wastewater to predict COVID-19 cases. Using a prospective experimental design, air, surface swabs, and wastewater samples were collected from a college dormitory housing roughly 500 students from March to May 2021 at the University of Miami, Coral Gables, FL. Students were randomly screened for COVID-19 during the study period. SARS-CoV-2 concentration in environmental samples was quantified using Volcano 2nd Generation-qPCR. Descriptive analyses were conducted to examine the associations between time-lagged SARS-CoV-2 in environmental samples and COVID-19 cases. SARS-CoV-2 was detected in air, surface swab and wastewater samples on 52 (63.4 %), 40 (50.0 %) and 57 (68.6 %) days, respectively. On 19 (24 %) of 78 days SARS-CoV-2 was detected in all three sample types. COVID-19 cases were reported on 11 days during the study period and SARS-CoV-2 was also detected two days before the case diagnosis on all 11 (100 %), 9 (81.8 %) and 8 (72.7 %) days in air, surface swab and wastewater samples, respectively. SARS-CoV-2 detection in environmental samples was an indicator of the presence of local COVID-19 cases and a 3-day lead indicator for a potential outbreak at the dormitory building scale. Proactive environmental surveillance of SARS-CoV-2 or other pathogens in multiple environmental media has potential to guide targeted measures to contain and/or mitigate infectious disease outbreaks within communities.


Assuntos
COVID-19 , Humanos , COVID-19/epidemiologia , SARS-CoV-2 , Águas Residuárias/análise , RNA Viral , Estudos Prospectivos
20.
Methods Mol Biol ; 2585: 127-143, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36331771

RESUMO

West Nile virus (WNV) is an important zoonotic pathogen, which is detected mainly by identification of its RNA using PCR. Genetic differentiation between WNV lineages is usually performed by complete genome sequencing, which is not available in many research and diagnostic laboratories. In this chapter, we describe a protocol for detection and analysis of WNV samples by sequencing the entire region of their structural genes capsid (C), preM/membrane, and envelope. The primary step is the detection of WNV RNA by quantitative PCR of the NS2A gene or the C gene regions. Next, the entire region containing the structural protein genes is amplified by PCR. The primary PCR product is then amplified again in parallel reactions, and these secondary PCR products are sequenced. Finally, bioinformatic analysis enables detection of mutations and classification of the samples of interest. This protocol is designed to be used by any laboratory equipped for endpoint and quantitative PCR. The sequencing can be performed either in-house or outsourced to a third-party service provider. This protocol may therefore be useful for rapid and affordable classification of WNV samples, obviating the need for complete genome sequencing.


Assuntos
Febre do Nilo Ocidental , Vírus do Nilo Ocidental , Animais , Vírus do Nilo Ocidental/genética , Febre do Nilo Ocidental/diagnóstico , Febre do Nilo Ocidental/genética , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Aves/genética , Proteínas Estruturais Virais/genética
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