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1.
Euro Surveill ; 28(28)2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37440348

RESUMO

After the near absence of influenza and other respiratory viruses during the first 2 years of the COVID-19 pandemic, an increased activity of mainly influenza A(H3N2) was detected at the beginning of August 2022 in Greece on three islands. Of 33 cases with respiratory symptoms testing negative for SARS-CoV-2 with rapid antigen tests, 24 were positive for influenza: 20 as A(H3N2) subtype and four as A(H1N1)pdm09 subtype. Phylogenetic analysis of selected samples from both subtypes was performed and they fell into clusters within subclades that included the 2022/23 vaccine strains. Our data suggest that influenza can be transmitted even in the presence of another highly infectious pathogen, such as SARS-CoV-2, with a similar transmission mode. We highlight the need for implementing changes in the current influenza surveillance and suggest a move from seasonal to continuous surveillance, especially in areas with a high number of tourists. Year-round surveillance would allow for a timelier start of vaccination campaigns and antiviral drugs procurement processes.


Assuntos
COVID-19 , Vírus da Influenza A Subtipo H1N1 , Vacinas contra Influenza , Influenza Humana , Humanos , Influenza Humana/diagnóstico , Influenza Humana/epidemiologia , Influenza Humana/prevenção & controle , Vírus da Influenza A Subtipo H3N2 , Grécia/epidemiologia , Filogenia , Pandemias , COVID-19/epidemiologia , Estações do Ano , SARS-CoV-2
2.
Molecules ; 27(13)2022 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-35807306

RESUMO

Ethnopharmacology, through the description of the beneficial effects of plants, has provided an early framework for the therapeutic use of natural compounds. Natural products, either in their native form or after crude extraction of their active ingredients, have long been used by different populations and explored as invaluable sources for drug design. The transition from traditional ethnopharmacology to drug discovery has followed a straightforward path, assisted by the evolution of isolation and characterization methods, the increase in computational power, and the development of specific chemoinformatic methods. The deriving extensive exploitation of the natural product chemical space has led to the discovery of novel compounds with pharmaceutical properties, although this was not followed by an analogous increase in novel drugs. In this work, we discuss the evolution of ideas and methods, from traditional ethnopharmacology to in silico drug discovery, applied to natural products. We point out that, in the past, the starting point was the plant itself, identified by sustained ethnopharmacological research, with the active compound deriving after extensive analysis and testing. In contrast, in recent years, the active substance has been pinpointed by computational methods (in silico docking and molecular dynamics, network pharmacology), followed by the identification of the plant(s) containing the active ingredient, identified by existing or putative ethnopharmacological information. We further stress the potential pitfalls of recent in silico methods and discuss the absolute need for in vitro and in vivo validation as an absolute requirement. Finally, we present our contribution to natural products' drug discovery by discussing specific examples, applying the whole continuum of this rapidly evolving field. In detail, we report the isolation of novel antiviral compounds, based on natural products active against influenza and SARS-CoV-2 and novel substances active on a specific GPCR, OXER1.


Assuntos
Produtos Biológicos , Tratamento Farmacológico da COVID-19 , Produtos Biológicos/química , Descoberta de Drogas/métodos , Etnofarmacologia/métodos , Plantas/química , SARS-CoV-2
3.
Molecules ; 26(19)2021 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-34641612

RESUMO

3CL-Pro is the SARS-CoV-2 main protease (MPro). It acts as a homodimer to cleave the large polyprotein 1ab transcript into proteins that are necessary for viral growth and replication. 3CL-Pro has been one of the most studied SARS-CoV-2 proteins and a main target of therapeutics. A number of drug candidates have been reported, including natural products. Here, we employ elaborate computational methods to explore the dimerization of the 3CL-Pro protein, and we formulate a computational context to identify potential inhibitors of this process. We report that fortunellin (acacetin 7-O-neohesperidoside), a natural flavonoid O-glycoside, and its structural analogs are potent inhibitors of 3CL-Pro dimerization, inhibiting viral plaque formation in vitro. We thus propose a novel basis for the search of pharmaceuticals as well as dietary supplements in the fight against SARS-CoV-2 and COVID-19.


Assuntos
Antivirais/farmacologia , Tratamento Farmacológico da COVID-19 , Proteases 3C de Coronavírus/antagonistas & inibidores , Flavonoides/farmacologia , Glicosídeos/farmacologia , Inibidores de Proteases/farmacologia , SARS-CoV-2/efeitos dos fármacos , Animais , Antivirais/química , Chlorocebus aethiops , Proteases 3C de Coronavírus/metabolismo , Flavonoides/química , Glicosídeos/química , Humanos , Simulação de Acoplamento Molecular , Polifenóis/química , Polifenóis/farmacologia , Inibidores de Proteases/química , Multimerização Proteica/efeitos dos fármacos , SARS-CoV-2/metabolismo , Células Vero
4.
Euro Surveill ; 25(32)2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32794446

RESUMO

BackgroundHuman cases of West Nile virus (WNV) infection are recorded since 2010 in Greece, with seasonal outbreaks occurring almost annually. Enhanced surveillance has been implemented since 2010, to promptly characterise cases' temporal and geographical distribution and inform authorities for implementation of appropriate measures (mosquito control, health education, blood safety).AimWe describe the epidemiology of WNV human infections in Greece focusing on the 2018 season.MethodsThe National Public Health Organization advised physicians to test all suspect WNV infection cases and refer samples to reference laboratories. Laboratories notified diagnosed cases on a daily basis. Treating physicians, patients, and infected blood donors were interviewed within 48 hours after diagnosis and the probable infection location was identified. Hospitalised cases were followed up until discharge.ResultsA total of 317 autochthonous WNV infection cases were diagnosed in 2018. Among them, 243 cases had neuroinvasive disease (WNND), representing a 23% increase of WNND cases compared with 2010, the previous most intense season. There were 51 deaths. Cases started occurring from week 22, earlier than usual. Both rural and urban areas were affected, with 86 (26% of the total) municipalities belonging to seven (54% of the total) regions recording cases. Two major epicentres were identified in Attica and Central Macedonia regions.ConclusionsThe largest number of human cases of WNV infection ever recorded in Greece occurred in 2018, with a wide geographical distribution, suggesting intense virus circulation. Enhanced surveillance is vital for the early detection of human cases and the prompt implementation of response measures.


Assuntos
Surtos de Doenças , Vigilância da População/métodos , Febre do Nilo Ocidental/epidemiologia , Vírus do Nilo Ocidental/isolamento & purificação , Animais , Anticorpos Antivirais/sangue , Doadores de Sangue , Feminino , Grécia/epidemiologia , Humanos , Estações do Ano , Febre do Nilo Ocidental/diagnóstico , Vírus do Nilo Ocidental/imunologia , Adulto Jovem
5.
Clin Exp Rheumatol ; 36(1): 102-109, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-28850029

RESUMO

OBJECTIVES: The risk of hepatitis B virus (HBV) reactivation with non-tumour necrosis factor inhibitor (non-TNFi) biologic agents in patients with rheumatic diseases and past HBV infection has not been definively elucidated. We assessed the comparative safety of non-TNFi and TNFi biologic agents in such patients in real-life clinical settings. METGODS: We carried out a retrospective cohort study from the Department of Rheumatology, University Hospital of Heraklion. Patients who received abatacept (ABA), tocilizumab (TCZ) or rituximab (RTX) during the period 2003-2016 and were HbsAg(-), anti-HBc(+), anti-HBs(±) at baseline, were monitored for HBV reactivation. Patients treated with TNFi agents during the same period were used as a control group. RESULTS: 101 cases of non-TNFi (39 ABA, 32 RTX and 30 TCZ) and 111 cases of TNFi treatment were identified. In non-TNFi, 76 cases (75.2%) were anti-HBc(+)/anti-HBs(+) and 25 (24.8%) were anti-HBc(+)/anti-HBs(-), as compared to 82 (73.9%) and 29 (26.1%) in TNFi-treated, respectively. After a median (IQR) observation of 24.0 (34.7) months, two cases (2.0%) of HBV reactivation were identified in the non-TNFi group; one with ABA, successfully treated with entecavir, and one fatal case with RTX and prior exposure to cyclophosphamide. No reactivation was observed in the TNFi group (p=0.226 vs. non-TNFi). Αnti-HBs titres were significantly reduced compared to baseline in the non-TNFi group [median (IQR) 203.9 (954.7) mIU/ml before treatment versus 144.9 (962.9) mIU/ml after treatment, p=0.03]. CONCLUSIONS: Two cases of HBV reactivation highlight the risk for this complication in patients with past HBV infection under biologic therapy.


Assuntos
Antirreumáticos/efeitos adversos , Artrite Reumatoide/tratamento farmacológico , Produtos Biológicos/efeitos adversos , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Vírus da Hepatite B/patogenicidade , Hepatite B/virologia , Ativação Viral , Adulto , Idoso , Artrite Reumatoide/diagnóstico , Artrite Reumatoide/imunologia , Doença Hepática Induzida por Substâncias e Drogas/diagnóstico , Feminino , Grécia , Hepatite B/diagnóstico , Hepatite B/imunologia , Vírus da Hepatite B/imunologia , Humanos , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/imunologia
6.
Graefes Arch Clin Exp Ophthalmol ; 256(1): 193-198, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29124395

RESUMO

PURPOSE: The evaluation of a non-invasive detection method for human papilloma virus (HPV) in ophthalmic pterygia. METHODS: Cotton swab samples and corresponding tissue specimens were collected from 21 ophthalmic pterygia of 21 patients. HPV detection and typing were performed by real-time PCR. Discrepancies in HPV detection between swab and tissue samples as well as clinical correlations of findings were examined. RESULTS: HPV DNA was detected in 9 (42.86%) tissue specimens and 8 (38.09%) respective swab specimens. HPV genotypes 33, 39, 45, 56, 59 and 66 were identified in the examined specimens, while more than one strain's HPV type was detected in 2 specimens. HPV presence was significantly correlated with the female gender whereas other clinical associations were not statistically significant. CONCLUSIONS: Findings imply that PCR-mediated HPV detection and typing in exfoliative swab specimens may be employed as a non-invasive diagnostic tool in the management of ophthalmic pterygia.


Assuntos
Túnica Conjuntiva/virologia , DNA Viral/análise , Infecções Oculares Virais/virologia , Papillomaviridae/genética , Infecções por Papillomavirus/virologia , Pterígio/complicações , Adulto , Idoso , Idoso de 80 Anos ou mais , Infecções Oculares Virais/complicações , Infecções Oculares Virais/diagnóstico , Estudos de Viabilidade , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/diagnóstico , Estudos Prospectivos , Pterígio/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real
7.
Cell Physiol Biochem ; 38(1): 94-109, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26741994

RESUMO

BACKGROUND/AIMS: Rho GTPases are crucial regulators of the actin cytoskeleton, membrane trafficking and cell signaling and their importance in cell migration and invasion is well- established. The human cytomegalovirus (HCMV) is a widespread pathogen responsible for generally asymptomatic and persistent infections in healthy people. Recent evidence indicates that HCMV gene products are expressed in over 90% of malignant type glioblastomas (GBM). In addition, the HCMV Immediate Early-1 protein (IE1) is expressed in >90% of tumors analyzed. METHODS: RhoA, RhoB and RhoC were individually depleted in U373MG glioblastoma cells as well as U373MG cells stably expressing the HCMV IE1 protein (named U373MG-IE1 cells) shRNA lentivirus vectors. Cell proliferation assays, migration as well as wound-healing assays were performed in uninfected and HCMV-infected cells. RESULTS: The depletion of RhoA, RhoB and RhoC protein resulted in significant alterations in the morphology of the uninfected cells, which were further enhanced by the cytopathic effect caused by HCMV. Furthermore, in the absence or presence of HCMV, the knockdown of RhoB and RhoC proteins decreased the proliferation rate of the parental and the IE1-expressing glioblastoma cells, whereas the knockdown of RhoA protein in the HCMV infected cell lines restored their proliferation rate. In addition, wound healing assays in U373MG cells revealed that depletion of RhoA, RhoB and RhoC differentially reduced their migration rate, even in the presence or the absence of HCMV. CONCLUSION: Collectively, these data show for the first time a differential implication of Rho GTPases in morphology, proliferation rate and motility of human glioblastoma cells during HCMV infection, further supporting an oncomodulatory role of HCMV depending on the Rho isoforms' state.


Assuntos
Citomegalovirus/fisiologia , Proteínas rho de Ligação ao GTP/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismo , Proteína rhoB de Ligação ao GTP/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Citomegalovirus/metabolismo , Glioblastoma/metabolismo , Glioblastoma/patologia , Células HEK293 , Humanos , Proteínas Imediatamente Precoces/genética , Proteínas Imediatamente Precoces/metabolismo , Microscopia de Fluorescência , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Imagem com Lapso de Tempo , Proteínas rho de Ligação ao GTP/antagonistas & inibidores , Proteínas rho de Ligação ao GTP/genética , Proteína rhoA de Ligação ao GTP/antagonistas & inibidores , Proteína rhoA de Ligação ao GTP/genética , Proteína rhoB de Ligação ao GTP/antagonistas & inibidores , Proteína rhoB de Ligação ao GTP/genética , Proteína de Ligação a GTP rhoC
8.
PLoS Pathog ; 10(5): e1004136, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24830456

RESUMO

Earlier studies had suggested that epigenetic mechanisms play an important role in the control of human cytomegalovirus (HCMV) infection. Here we show that productive HCMV infection is indeed under the control of histone H3K27 trimethylation. The histone H3K27 methyltransferase EZH2, and its regulators JARID2 and NDY1/KDM2B repress GFI1, a transcriptional repressor of the major immediate-early promoter (MIEP) of HCMV. Knocking down EZH2, NDY1/KDM2B or JARID2 relieves the repression and results in the upregulation of GFI1. During infection, the incoming HCMV rapidly downregulates the GFI1 mRNA and protein in both wild-type cells and in cells in which EZH2, NDY1/KDM2B or JARID2 were knocked down. However, since the pre-infection levels of GFI1 in the latter cells are significantly higher, the virus fails to downregulate it to levels permissive for MIEP activation and viral infection. Following the EZH2-NDY1/KDM2B-JARID2-independent downregulation of GFI1 in the early stages of infection, the virus also initiates an EZH2-NDY1/ΚDM2Β-JARID2-dependent program that represses GFI1 throughout the infection cycle. The EZH2 knockdown also delays histone H3K27 trimethylation in the immediate early region of HCMV, which is accompanied by a drop in H3K4 trimethylation that may contribute to the shEZH2-mediated repression of the major immediate early HCMV promoter. These data show that HCMV uses multiple mechanisms to allow the activation of the HCMV MIEP and to prevent cellular mechanisms from blocking the HCMV replication program.


Assuntos
Infecções por Citomegalovirus , Citomegalovirus/fisiologia , Proteínas de Ligação a DNA/genética , Proteínas F-Box/fisiologia , Histona Desmetilases com o Domínio Jumonji/fisiologia , Complexo Repressor Polycomb 2/fisiologia , Regiões Promotoras Genéticas , Fatores de Transcrição/genética , Antígenos Virais/genética , Células Cultivadas , Citomegalovirus/genética , Infecções por Citomegalovirus/genética , Infecções por Citomegalovirus/virologia , Proteínas de Ligação a DNA/metabolismo , Regulação para Baixo/genética , Proteína Potenciadora do Homólogo 2 de Zeste , Células HEK293 , Células HeLa , Humanos , Proteínas Imediatamente Precoces/genética , Transdução de Sinais/fisiologia , Fatores de Transcrição/metabolismo , Proteínas Virais/genética , Proteínas Virais/fisiologia , Replicação Viral/genética
9.
Arch Virol ; 160(4): 887-92, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25666196

RESUMO

Merkel cell polyomavirus (MCPyV) is a newly discovered human small, non-enveloped, double-stranded DNA virus, which was classified into the Polyomaviridae family. MCPyV is acquired in early childhood through close contact involving respiratory tract secretions and causes a widespread, previously unrecognised, asymptomatic infection in both immunocompetent children and adults. To date, several researchers have established that MCPyV is the potential causative agent of Merkel cell carcinoma, a relatively rare but life-threatening skin cancer of neuroendocrine origin. In our review, we present current data on the presence of MCPyV DNA in children and address the possible role that the respiratory tract plays in the route of viral transmission. Future studies are required to fully elucidate the potential implications of MCPyV infection in children.


Assuntos
Carcinoma de Célula de Merkel/virologia , Poliomavírus das Células de Merkel/fisiologia , Infecções por Polyomavirus/virologia , Infecções Tumorais por Vírus/virologia , Adolescente , Adulto , Carcinoma de Célula de Merkel/epidemiologia , Criança , Pré-Escolar , Humanos , Poliomavírus das Células de Merkel/genética , Infecções por Polyomavirus/epidemiologia , Infecções Tumorais por Vírus/epidemiologia , Adulto Jovem
10.
Proc Natl Acad Sci U S A ; 109(10): E613-21, 2012 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-22315412

RESUMO

The protein kinases Akt1, Akt2, and Akt3 possess nonredundant signaling properties, few of which have been investigated. Here, we present evidence for an Akt1-dependent pathway that controls interferon (IFN)-regulated gene expression and antiviral immunity. The target of this pathway is EMSY, an oncogenic interacting partner of BRCA2 that functions as a transcriptional repressor. Overexpression of EMSY in hTERT-immortalized mammary epithelial cells, and in breast and ovarian carcinoma cell lines, represses IFN-stimulated genes (ISGs) in a BRCA2-dependent manner, whereas its knockdown has the opposite effect. EMSY binds to the promoters of ISGs, suggesting that EMSY functions as a direct transcriptional repressor. Akt1, but not Akt2, phosphorylates EMSY at Ser209, relieving EMSY-mediated ISG repression. The Akt1/EMSY/ISG pathway is activated by both viral infection and IFN, and it inhibits the replication of HSV-1 and vesicular stomatitis virus (VSV). Collectively, these data define an Akt1-dependent pathway that contributes to the full activation of ISGs by relieving their repression by EMSY and BRCA2.


Assuntos
Proteínas de Neoplasias/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Repressoras/metabolismo , Células 3T3 , Animais , Proteína BRCA2/metabolismo , Linhagem Celular Tumoral , Humanos , Interferons/metabolismo , Camundongos , Camundongos Knockout , Modelos Biológicos , Oxirredutases atuantes sobre Doadores de Grupo CH-CH , Proteínas/metabolismo , Transcrição Gênica
11.
Tumour Biol ; 35(12): 12765-73, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25213701

RESUMO

Prostate cancer is the most common neoplasm found in males and the second most frequent cause of cancer-related mortality in males in Greece. Among other pathogens, the detection frequency of human papillomavirus (HPV) has been found to be significantly increased in tumor tissues among patients with sexually transmitted diseases (STDs), depending on the geographical distribution of each population studied. The present study focused on the detection of HPV and the distribution of Arg72Pro p53 polymorphism in a cohort of healthy individuals, as well as prostate cancer patients. We investigated the presence of HPV in 50 paraffin-embedded prostate cancer tissues, as well as in 30 physiological tissue samples from healthy individuals by real-time PCR. Furthermore, the same group of patients was also screened for the presence of the Arg72Pro polymorphism of the p53 gene, a p53 polymorphism related to HPV. Out of the 30 control samples, only 1 was found positive for HPV (3.33 %). On the contrary, HPV DNA was detected in 8 out of the total 50 samples (16 %) in the prostate cancer samples. The distribution of the three genotypes, Arg/Arg, Arg/Pro, and Pro/Pro, was 69.6, 21.7, and 8.7 % in the cancer patients and 75.0, 17.86, and 7.14 % in healthy controls, respectively. No statistically significant association was observed between the HPV presence and the age, stage, p53 polymorphism status at codon 72, or PSA. The increased prevalence of HPV detected in the prostate cancer tissues is in agreement with that reported in previous studies, further supporting the association of HPV infection and prostate cancer.


Assuntos
Alphapapillomavirus/genética , Códon , Infecções por Papillomavirus/complicações , Polimorfismo Genético , Neoplasias da Próstata/genética , Neoplasias da Próstata/virologia , Proteína Supressora de Tumor p53/genética , Idoso , Substituição de Aminoácidos , Estudos de Casos e Controles , Grécia , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Neoplasias da Próstata/patologia
12.
Tumour Biol ; 35(4): 3203-9, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24310500

RESUMO

Although the role of human papillomavirus (HPV) in the development of uterine cervical cancer is well established, the role of HPV in lung carcinogenesis remains controversial. The detection rates of HPV DNA are subject to a wide variation from 0 to 100%. This is partly influenced by the detection techniques employed. To elucidate the impact of HPV infection on lung parenchyma, we analyzed 100 non-small cell lung cancer (NSCLC) specimens (39 squamous cell carcinomas, 50 adenocarcinomas, 5 samples with characteristics of both squamous cell and adenocarcinoma, 5 undifferentiated and 1 large cell carcinoma) from the region of Crete, Greece. Sixteen non-cancerous samples served as the negative controls. DNA was extracted from 100 paraffin-embedded tissue sections obtained from NSCLC patients. The specimens were examined for the detection of HPV DNA by Real-Time PCR using GP5+/GP6+ primers. Furthermore, the HPV-positive samples were subjected to genotyping. In contrast to the absence of viral genomes in the control samples, HPV DNA was detected in 19 NSCLC specimens (19%). In particular, 4 squamous cell carcinomas, 12 adenocarcinomas, 1 sample with characteristics of both squamous cell and adenocarcinoma, and 2 undifferentiated samples were HPV-positive. The distribution of HPV genotypes was as follows: HPV 16: eight cases (42.1%); HPV 11: three cases (15.8%); HPV 6: one case (5.2%); HPV 59: one case (5.2%); HPV 33: two cases (10.5%); HPV 31: two cases (10.5%) and HPV 18: two cases (10.5%). The presence of HPV in the tumor samples provides evidence of the potential role of HPV in NSCLC and strongly argues for additional research on this issue.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/virologia , Neoplasias Pulmonares/virologia , Papillomaviridae/isolamento & purificação , Idoso , Carcinoma Pulmonar de Células não Pequenas/fisiopatologia , DNA Viral/análise , Feminino , Volume Expiratório Forçado , Genótipo , Humanos , Neoplasias Pulmonares/fisiopatologia , Masculino , Pessoa de Meia-Idade , Papillomaviridae/genética , Capacidade Vital
13.
Microbiol Spectr ; : e0067224, 2024 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-39422469

RESUMO

The goal of this study is to test a novel device and methodology based on the "Pebble" platform and real-time quantitative colorimetric loop-mediated isothermal amplification (qcLAMP) during SARS-CoV-2 detection using crude samples and extracted RNA. The new method employs an inexpensive lightweight device aimed toward rapid point-of-care testing. An extensive evaluation was performed consisting of 1,693 clinical samples across five independent clinical testing centers. Positive colorimetric results were observed within 20 minutes of testing. At a 20-minute time-to-positive cut-off, the specificity is 98.5% with a diagnostic accuracy of 91.9%, compared to qPCR assays. Our findings indicate that the SARS-CoV-2 qcLAMP diagnostic assay in conjunction with the Pebble device is ideal for point-of-care/near-patient testing.IMPORTANCEHere, we describe our analyses and validation of a novel real-time quantitative colorimetric loop-mediated isothermal amplification (qcLAMP) device, available under the name "Pebble" and associated SARS-CoV-2 diagnostic qcLAMP assay for clinical diagnostic use. The analyses were performed in five independent testing sites across Europe using clinical samples from the associated clinical sites and support the use of "pebble" and associated kit in the diagnostic environment.

15.
Lancet Reg Health Eur ; 40: 100892, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38590940

RESUMO

Congenital cytomegalovirus (cCMV) infection carries a significant burden with a 0.64% global prevalence and a 17-20% chance of serious long-term effects in children. Since the last guidelines, our understanding, particularly regarding primary maternal infections, has improved. A cCMV guidelines group was convened under the patronage of the European Society of Clinical Virology in April 2023 to refine these insights. The quality and validity of selected studies were assessed for potential biases and the GRADE framework was employed to evaluate quality of evidence across key domains. The resulting recommendations address managing cCMV, spanning prevention to postnatal care. Emphasizing early and accurate maternal diagnosis through serological tests enhances risk management and prevention strategies, including using valaciclovir to prevent vertical transmission. The guidelines also strive to refine personalized postnatal care based on risk assessments, ensuring targeted interventions for affected families.

16.
J Clin Virol Plus ; 3(1): 100129, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36530947

RESUMO

The SARS-CoV-2 pandemic led to an urgent need for rapid diagnostic testing in order to inform timely patients' management. This study aimed to assess the performance of the STANDARD™ M10 SARS-CoV-2 assay as a diagnostic tool for COVID-19. A total of 400 nasopharyngeal or oropharyngeal swabs were tested against a reference real-time RT-PCR, including 200 positive samples spanning the full range of observed Ct values. The sensitivity of the STANDARD™ M10 SARS-CoV-2 assay was 98.00% (95% CI 94.96% to 99.45%, 196/200), while the specificity was also estimated at 97.50% (95% CI 94.26% to 99.18%, 195/200). The assay proved highly efficient for the detection of SARS-CoV-2, even in samples with low viral load (Ct>25), presenting lower Ct values compared to the reference method. We concluded that the STANDARD™ M10 SARS-CoV-2 assay has a similar performance compared to the reference method and other molecular point-of-care assays and can be a valuable tool for rapid and accurate diagnosis.

17.
Infect Dis (Lond) ; 55(7): 509-513, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37198922

RESUMO

BACKGROUND: Rapid and accurate diagnostics of patients with suspected seasonal influenza or pathogens of the upper respiratory tract is crucial. Fast detection is important especially for influenza A/B virus, so that isolation measures should be taken to prevent the spread of the virus. METHODS: We compared the performance of two syndromic testing methodologies (QIAstat-Dx RP, BioFire RP2plus) against the Alere™ i as the comparator method. Totally, 97 swab samples were included from patients with symptoms of acute respiratory infection admitted in the hospitals of the wider region of Crete, Greece. RESULTS: The Positive Percent Agreement (PPA) of the BioFire RP2plus was 100% (95% CI 87.66%-100%), while the Negative Percent Agreement (NPA) was estimated at 91.3% (95% CI 82.03%-96.74%). This method produced no invalid results. For QIAstat-Dx RP the PPA was 89.29% (95% CI 71.77%-97.73%), while the NPA was 91.3% (95% CI 82.03%-96.74%, 63/69). The BioFire RP2plus managed to determine the subtype in more samples than the QIAstat-Dx RP. CONCLUSIONS: Both panels can be valuable tools for clinicians, since they both display high sensitivity and specificity. We report a slightly better performance for BioFire RP2plus, since it produced no invalid results.


Assuntos
Herpesvirus Cercopitecino 1 , Vírus da Influenza A , Influenza Humana , Humanos , Influenza Humana/diagnóstico , Vírus da Influenza B/genética , Vírus da Influenza A/genética , Sistemas Automatizados de Assistência Junto ao Leito , Técnicas de Diagnóstico Molecular/métodos , Sensibilidade e Especificidade
18.
Diseases ; 11(3)2023 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-37606476

RESUMO

INTRODUCTION: The need for effective therapeutic regimens for non-critically ill patients during the COVID-19 pandemic remained largely unmet. Previous work has shown that a combination of three aromatic plants' essential oils (CAPeo) (Thymbra capitata (L.) Cav., Origanum dictamnus L., Salvia fruticose Mill.) has remarkable in vitro antiviral activity. Given its properties, it was urgent to explore its potential in treating mild COVID-19 patients in primary care settings. METHODS: A total of 69 adult patients were included in a clinical proof-of-concept (PoC) intervention study. Family physicians implemented the observational study in two arms (intervention group and control group) during three study periods (IG2020, n=13, IG2021/22, n=25, and CG2021/22, n=31). The SARS-CoV-2 infection was confirmed by real-time PCR. The CAPeo mixture was administered daily for 14 days per os in the intervention group, while the control group received usual care. RESULTS: The PoC study found that the number and frequency of general symptoms, including general fatigue, weakness, fever, and myalgia, decreased following CAPeo administration. By Day 7, the average presence (number) of symptoms decreased in comparison with Day 1 in IG (4.7 to 1.4) as well as in CG (4.0 to 3.1), representing a significant decrease in the cumulative presence in IC (-3.3 vs. -0.9, p < 0.001; η2 = 0.20) on Day 7 and on Day 14 (-4.2 vs. -2.9, p = 0.027; η2 = 0.08). DISCUSSION/CONCLUSIONS: Our findings suggest that CAPeo possesses potent antiviral activity against SARS-CoV-2 in addition tο its effect against influenza A and B and human rhinovirus HRV14 strains. The early and effective impact on alleviating key symptoms of COVID-19 may suggest this mixture can act as a complementary natural agent for patients with mild COVID-19.

19.
J Virol Methods ; 319: 114759, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37257757

RESUMO

The circulation of certain SARS-CoV-2 variants may have a great impact on the epidemiological status of a geographical area; therefore, it is important that their presence is monitored. Currently, the gold standard method used to identify newly emerged variants is sequencing of either genes or whole genomes. However, since this method is relatively expensive and has a long turnaround time, other rapid strategies should also be employed. The current study aimed to evaluate the performance of the Simplexa® SARS-CoV-2 Variants Direct assay, which is a RT-PCR that determines the variant present in a nasopharyngeal swab sample in approximately two hours. Totally, 527 positive samples for SARS-CoV-2 were analyzed from January until December 2022 and next-generation sequencing (NGS) was used as the reference method. The assay showed high sensitivity, ranging from 94.12 % to 100 %, depending on the variant. The assay also showed high specificity, reaching 100 % for Delta and BA.1 variants, and 99.74 % and 98.67 % for BA.2 and BA.4/BA.5 variants, respectively. Moreover, the assay was able to identify the correct variant category in the presence of any subvariant in the sample. We conclude that the assay can be used to facilitate faster monitoring of circulating SARS-CoV-2 variants, however sequencing cannot be completely replaced, since new variants always emerge, and constant updates are needed, so that the user is able to interpret the melting curve patterns.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , COVID-19/diagnóstico , Bioensaio , Sequenciamento de Nucleotídeos em Larga Escala
20.
Viruses ; 15(3)2023 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-36992465

RESUMO

Patients receiving treatment with B-cell-depleting monoclonal antibodies, such as anti-CD20 monoclonal antibodies, such as rituximab and obinutuzumab, either for hematological disease or another diagnosis, such as a rheumatological disease, are at an increased risk for medical complications and mortality from COVID-19. Since inconsistencies persist regarding the use of convalescent plasma (CP), especially in the vulnerable patient population that has received previous treatment with B-cell-depleting monoclonal antibodies, further studies should be performed in thisdirection. The aim of the present study was to describe the characteristics of patients with previous use of B-cell-depleting monoclonal antibodies and describe the potential beneficial effects of CP use in terms of mortality, ICU admission and disease relapse. In this retrospective cohort study, 39 patients with previous use of B-cell-depleting monoclonal antibodies hospitalized in the COVID-19 department of a tertiary hospital in Greece were recorded and evaluated. The mean age was 66.3 years and 51.3% were male. Regarding treatment for COVID-19, remdesivir was used in 89.7%, corticosteroids in 94.9% and CP in 53.8%. In-hospital mortality was 15.4%. Patients who died were more likely to need ICU admission and also had a trend towards a longer hospital stay, even though the last did not reach statistical significance. Patients treated with CP had a lower re-admission rate for COVID-19 after discharge. Further studies should be performed to identify the role of CP in patients with treatment with B-cell-depleting monoclonal antibodies suffering from COVID-19.


Assuntos
COVID-19 , Humanos , Masculino , Idoso , Feminino , COVID-19/terapia , COVID-19/etiologia , SARS-CoV-2 , Estudos Retrospectivos , Imunização Passiva/efeitos adversos , Soroterapia para COVID-19 , Anticorpos Monoclonais/uso terapêutico
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