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BACKGROUND: Pandemic coronavirus disease 2019 (COVID-19) disease represents a challenge for healthcare structures. The molecular confirmation of samples from infected individuals is crucial and therefore guides public health decision making. Clusters and possibly increased diffuse transmission could occur in the context of the next influenza season. For this reason, a diagnostic test able to discriminate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from influenza viruses is urgently needed. METHODS: A multiplex real-time reverse-transcription polymerase chain reaction (PCR) assay was assessed using 1 laboratory protocol with different real-time PCR instruments. Overall, 1000 clinical samples (600 from samples SARS-CoV-2-infected patients, 200 samples from influenza-infected patients, and 200 negative samples) were analyzed. RESULTS: The assay developed was able to detect and discriminate each virus target and to intercept coinfections. The limit of quantification of each assay ranged between 5 and 10 genomic copy numbers, with a cutoff value of 37.7 and 37.8 for influenza and SARS-CoV-2 viruses, respectively. Only 2 influenza coinfections were detected in COVID-19 samples. CONCLUSIONS: This study suggests that multiplex assay is a rapid, valid, and accurate method for the detection of SARS-CoV-2 and influenza viruses in clinical samples. The test may be an important diagnostic tool for both diagnostic and surveillance purposes during the seasonal influenza activity period.
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COVID-19/diagnóstico , Influenza Humana/diagnóstico , Orthomyxoviridae/isolamento & purificação , SARS-CoV-2/isolamento & purificação , Área Sob a Curva , COVID-19/complicações , COVID-19/epidemiologia , Diagnóstico Diferencial , Humanos , Influenza Humana/complicações , Influenza Humana/epidemiologia , Reação em Cadeia da Polimerase Multiplex , Orthomyxoviridae/genética , RNA Viral/isolamento & purificação , Curva ROC , Reprodutibilidade dos Testes , SARS-CoV-2/genética , Estações do Ano , Sensibilidade e EspecificidadeRESUMO
Whole genome sequences of SARS-CoV-2 obtained from two patients, a Chinese tourist visiting Rome and an Italian, were compared with sequences from Europe and elsewhere. In a phylogenetic tree, the Italian patient's sequence clustered with sequences from Germany while the tourist's sequence clustered with other European sequences. Some additional European sequences in the tree segregated outside the two clusters containing the patients' sequences. This suggests multiple SARS-CoV-2 introductions in Europe or virus evolution during circulation.
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Betacoronavirus/genética , Infecções por Coronavirus/diagnóstico , Coronavirus/genética , Genoma Viral/genética , Pneumonia Viral/diagnóstico , RNA Viral/genética , Síndrome Respiratória Aguda Grave/diagnóstico , Viagem , Sequenciamento Completo do Genoma/métodos , Betacoronavirus/isolamento & purificação , COVID-19 , China , Coronavirus/classificação , Coronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/transmissão , Alemanha , Humanos , Itália , Epidemiologia Molecular , Pandemias , Filogenia , Pneumonia Viral/epidemiologia , Pneumonia Viral/transmissão , Mutação Puntual , RNA Viral/isolamento & purificação , SARS-CoV-2 , Síndrome Respiratória Aguda Grave/virologiaRESUMO
BackgroundOn 20 February 2020, a locally acquired coronavirus disease (COVID-19) case was detected in Lombardy, Italy. This was the first signal of ongoing transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the country. The number of cases in Italy increased rapidly and the country became the first in Europe to experience a SARS-CoV-2 outbreak.AimOur aim was to describe the epidemiology and transmission dynamics of the first COVID-19 cases in Italy amid ongoing control measures.MethodsWe analysed all RT-PCR-confirmed COVID-19 cases reported to the national integrated surveillance system until 31 March 2020. We provide a descriptive epidemiological summary and estimate the basic and net reproductive numbers by region.ResultsOf the 98,716 cases of COVID-19 analysed, 9,512 were healthcare workers. Of the 10,943 reported COVID-19-associated deaths (crude case fatality ratio: 11.1%) 49.5% occurred in cases older than 80 years. Male sex and age were independent risk factors for COVID-19 death. Estimates of R0 varied between 2.50 (95% confidence interval (CI): 2.18-2.83) in Tuscany and 3.00 (95% CI: 2.68-3.33) in Lazio. The net reproduction number Rt in northern regions started decreasing immediately after the first detection.ConclusionThe COVID-19 outbreak in Italy showed a clustering onset similar to the one in Wuhan, China. R0 at 2.96 in Lombardy combined with delayed detection explains the high case load and rapid geographical spread. Overall, Rt in Italian regions showed early signs of decrease, with large diversity in incidence, supporting the importance of combined non-pharmacological control measures.
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Número Básico de Reprodução , COVID-19/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , COVID-19/mortalidade , COVID-19/transmissão , Feminino , Pessoal de Saúde/estatística & dados numéricos , Humanos , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Mortalidade , SARS-CoV-2RESUMO
OBJECTIVES: to describe the integrated surveillance system of COVID-19 in Italy, to illustrate the outputs used to return epidemiological information on the spread of the epidemic to the competent public health bodies and to the Italian population, and to describe how the surveillance data contributes to the ongoing weekly regional monitoring and risk assessment system. METHODS: the COVID-19 integrated surveillance system is the result of a close and continuous collaboration between the Italian National Institute of Health (ISS), the Italian Ministry of Health, and the regional and local health authorities. Through a web platform, it collects individual data of laboratory confirmed cases of SARS-CoV-2 infection and gathers information on their residence, laboratory diagnosis, hospitalisation, clinical status, risk factors, and outcome. Results, for different levels of aggregation and risk categories, are published daily and weekly on the ISS website, and made available to national and regional public health authorities; these results contribute one of the information sources of the regional monitoring and risk assessment system. RESULTS: the COVID-19 integrated surveillance system monitors the space-time distribution of cases and their characteristics. Indicators used in the weekly regional monitoring and risk assessment system include process indicators on completeness and results indicators on weekly trends of newly diagnosed cases per Region. CONCLUSIONS: the outputs of the integrated surveillance system for COVID-19 provide timely information to health authorities and to the general population on the evolution of the epidemic in Italy. They also contribute to the continuous re-assessment of risk related to transmission and impact of the epidemic thus contributing to the management of COVID-19 in Italy.
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COVID-19/epidemiologia , Vigilância da População , SARS-CoV-2 , Hospitalização/estatística & dados numéricos , Humanos , Disseminação de Informação , Itália/epidemiologia , Vigilância da População/métodos , Relatório de Pesquisa , RiscoRESUMO
Both malaria and relapsing fever Borrelia are infectious diseases characterized by fever, headache, myalgia, hepatosplenomegaly and tendency to relapse. Exflagellation of microgametocyte in malarial parasites is seen only in the definitive host, i.e., mosquitoes. Here we report an unusual case of a 23-year-old man who presented Plasmodium vivax infection with multiple exflagellated microgametes in the peripheral blood smear.
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Malária Vivax , Plasmodium vivax , Adulto , Animais , Cloroquina/uso terapêutico , Humanos , Estágios do Ciclo de Vida , Malária Vivax/sangue , Malária Vivax/diagnóstico , Malária Vivax/tratamento farmacológico , Masculino , Plasmodium vivax/crescimento & desenvolvimento , Recidiva , Resultado do Tratamento , Adulto JovemRESUMO
OBJECTIVE: To describe a rare case of acute Q fever with tache noire. CLINICAL PRESENTATION AND INTERVENTION: A 51-year-old man experienced acute Q fever showing tache noire, generally considered a pathognomonic sign of Mediterranean spotted fever (MSF) and MSF-like illness, but not a clinical feature of Q fever. The patient was treated with doxycycline 100 mg every 12 h. CONCLUSION: In the Mediterranean area, tache noire should be considered pathognomonic of MSF but it should not rule out Q fever. Clinical diagnosis should be supported by accurate laboratory diagnostic tests to guide proper management.
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Febre Q/diagnóstico , Febre Q/fisiopatologia , Febre Botonosa/diagnóstico , Febre Botonosa/fisiopatologia , Diagnóstico Diferencial , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
A serious respiratory tract infection due to Corynebacterium ulcerans was observed in a 70-year-old woman. Clinical presentation included pseudomembranes in the upper respiratory tract and lung involvement. C. ulcerans was recovered from the nose of the patient's dog. Both dog's and patient's isolates belonged to Sequence Type 331.
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Difteria/diagnóstico , Infecções Respiratórias/diagnóstico , Idoso , Animais , Difteria/tratamento farmacológico , Difteria/microbiologia , Doenças do Cão/diagnóstico , Doenças do Cão/tratamento farmacológico , Doenças do Cão/microbiologia , Doenças do Cão/transmissão , Cães , Feminino , Humanos , Itália , Infecções Respiratórias/tratamento farmacológico , Infecções Respiratórias/microbiologia , Zoonoses/diagnóstico , Zoonoses/tratamento farmacológico , Zoonoses/microbiologiaRESUMO
INTRODUCTION: Rickettsioses represent a group of emerging infectious diseases in Europe. Climate changes and the anthropization of rural environment have favored vectors' biological cycle and geographic spread. In Sardinia, Mediterranean spotted fever (MSF) is endemic and represents an important public health problem. PURPOSE: We investigated the etiology and the clinical presentation of MSF-like illness in northern Sardinia by enrolling patients admitted to the Infectious Disease Unit of the University of Sassari. RESULTS: Diagnostic tests included ELISA, Indirect immunofluorescence (IFI), DNA isolation from blood and from eschar samples with real-time PCR and genotyping. Eighty-seven patients with a mean age of 53 ± 14 years, of whom 65 (75 %) males, were included in the study. The most common diagnosis was MSF (79 %), followed by Q fever (8 %), and anaplasmosis (2 %). A tache noire was found in 58 % of rickettioses and 28 % of Coxiella burnetii infections. MSF was confirmed in 47 % of the cases by IFI and 43 % by ELISA antibody tests. The isolation of rickettsial DNA from the eschar was positive in 10/13 (77 %) of the cases due to Rickettsia conorii. Using this method, we identified the first case of R. monacensis infection in Italy. CONCLUSIONS: In conclusion, antibody-based tests confirmed the diagnosis in less than 50 % of the cases, whereas DNA isolation confirmed the diagnosis in 77 % of tested cases and allowed the identification of a new pathogenic species in Italy. Therefore, DNA isolation should be implemented to better identify the etiology of MSF-like illnesses and help the clinician in the management of patients.
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Febre Botonosa/diagnóstico , Febre Botonosa/microbiologia , Rickettsia conorii/genética , Adulto , Idoso , Febre Botonosa/epidemiologia , Febre Botonosa/fisiopatologia , DNA Bacteriano/genética , Feminino , Humanos , Itália , Masculino , Pessoa de Meia-IdadeRESUMO
We report the first human fatal case of streptococcal toxic shock syndrome (STSS) caused by Streptococcus suis serotype 2 carrying the tetracycline efflux tet (40) gene and the tetracycline ribosomal protection tet (O/W/32/O) gene. The patient was splenectomized. The case was characterized by multi-organ dysfunction and disseminated intravascular coagulopathy, in accordance with the clinical parameters of STSS. More investigations are needed to improve the epidemiology and the pathogenesis of S. suis in human infection.
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Choque Séptico/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus suis/genética , Proteínas de Bactérias/genética , Humanos , Itália , Masculino , Pessoa de Meia-IdadeRESUMO
Phospholipases are produced from bacterial pathogens causing very different diseases. One of the most intriguing aspects of phospholipases is their potential to interfere with cellular signaling cascades and to modulate the host-immune response. Here, we investigated the role of the innate and acquired immune responses elicited by Chlamydophila pneumoniae phospholipase D (CpPLD) in the pathogenesis of atherosclerosis. We evaluated the cytokine and chemokine production induced by CpPLD in healthy donors' monocytes and in vivo activated T cells specific for CpPLD that infiltrate atherosclerotic lesions of patients with C. pneumoniae antibodies. We also examined the helper function of CpPLD-specific T cells for monocyte matrix metalloproteinase (MMP)-9 and tissue factor (TF) production as well as the CpPLD-induced chemokine expression by human venular endothelial cells (HUVECs). We report here that CpPLD is a TLR4 agonist able to induce the expression of IL-23, IL-6, IL-1ß, TGF-ß, and CCL-20 in monocytes, as well as CXCL-9, CCL-20, CCL-4, CCL-2, ICAM-1, and VCAM-1 in HUVECs. Plaque-derived T cells produce IL-17 in response to CpPLD. Moreover, CpPLD-specific CD4(+) T lymphocytes display helper function for monocyte MMP-9 and TF production. CpPLD promotes Th17 cell migration through the induction of chemokine secretion and adhesion molecule expression on endothelial cells. These findings indicate that CpPLD is able to drive the expression of IL-23, IL-6, IL-1ß, TGF-ß, and CCL-20 by monocytes and to elicit a Th17 immune response that plays a key role in the genesis of atherosclerosis.
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Aterosclerose/imunologia , Aterosclerose/microbiologia , Chlamydophila pneumoniae/enzimologia , Regulação da Expressão Gênica/imunologia , Fosfolipase D/imunologia , Células Th17/imunologia , Idoso , Linhagem Celular , Quimiocinas/imunologia , Citocinas/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Metaloproteinase 9 da Matriz/metabolismo , Pessoa de Meia-Idade , Monócitos/imunologia , Fosfolipase D/farmacologia , Reação em Cadeia da Polimerase em Tempo Real , Tromboplastina/metabolismo , Receptor 4 Toll-Like/agonistasRESUMO
Chlamydophila pneumoniae, an aetiological agent of respiratory infection, is also thought to play an immuno-pathogenetic role in atherosclerosis by contributing to inflammation and plaque instability. Phospholipase D (PLD) is an enzyme involved in lipid metabolism and may have a direct or indirect impact on virulence and the inflammatory response. Some aspects of the developmental cycle of C. pneumoniae suggest a direct implication of its PLD (CpPLD) in the pathogenesis, specifically by affecting the regulation of lipid metabolism and lipid exchange between C. pneumoniae and host cells. Our previous studies disclosed a specific anti-CpPLD antibody response in patients with acute coronary syndromes chronically infected with C. pneumoniae, and demonstrated that this antigen is a factor able to drive the inflammatory process in atherosclerosis. Due to the intriguing aspects of the CpPLD, the present study investigated CpPLD enzymatic activity of the protein and the two domains that include one HKD motif each polypeptide. Our results showed that CpPLD was able to synthesize the cardiolipin (CL) but unable to hydrolyze phospholipids. It was also observed that each single HKD motif has an independent CL synthetase activity. This enzymatic activity of CpPLD could be important in the inflammatory process within the atherothrombotic events.
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Proteínas de Bactérias/metabolismo , Infecções por Chlamydophila/microbiologia , Chlamydophila pneumoniae/enzimologia , Fosfolipase D/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Cardiolipinas/metabolismo , Chlamydophila pneumoniae/química , Chlamydophila pneumoniae/genética , Humanos , Hidrólise , Dados de Sequência Molecular , Fosfolipase D/química , Fosfolipase D/genética , Fosfolipídeos/metabolismo , Estrutura Terciária de ProteínaAssuntos
Imunodeficiência de Variável Comum , Infecções por Haemophilus , Haemophilus influenzae , Infecções Pneumocócicas , Infecções Respiratórias , Streptococcus pneumoniae , Adolescente , Adulto , Idoso , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Imunodeficiência de Variável Comum/imunologia , Imunodeficiência de Variável Comum/microbiologia , Feminino , Infecções por Haemophilus/imunologia , Infecções por Haemophilus/microbiologia , Haemophilus influenzae/efeitos dos fármacos , Haemophilus influenzae/genética , Haemophilus influenzae/crescimento & desenvolvimento , Haemophilus influenzae/isolamento & purificação , Humanos , Imunoglobulina A/sangue , Imunoglobulina G/uso terapêutico , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Nasofaringe/microbiologia , Orofaringe/microbiologia , Infecções Pneumocócicas/imunologia , Infecções Pneumocócicas/microbiologia , Infecções Respiratórias/imunologia , Infecções Respiratórias/microbiologia , Fatores de Risco , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/crescimento & desenvolvimento , Streptococcus pneumoniae/isolamento & purificação , Adulto JovemRESUMO
The role of migratory birds in circulation tick-borne viruses needs to be better defined. In order to assess the potential role of migratory birds in exotic virus spread, we conducted a study to identify ticks collected from migratory birds in the Central Region of Italy, and performed molecular investigation for Crimea-Congo Hemorrhagic Fever (CCHFV), West Nile fever (WNFV) and Usutu (USUV) in the vectors. A total of 137 competent ticks were collected with predominance of Hyalomma species. Although, negative results were obtained for all viruses considered, the high proportion of Hyalomma ticks highlights the potential risk for the dissemination of tick-borne viruses through infested migratory birds.
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Vetores Aracnídeos/virologia , Aves/parasitologia , Vírus da Febre Hemorrágica da Crimeia-Congo/isolamento & purificação , Carrapatos/virologia , Vírus do Nilo Ocidental/isolamento & purificação , Migração Animal , Animais , Vetores Aracnídeos/classificação , Aves/fisiologia , Vírus da Febre Hemorrágica da Crimeia-Congo/genética , Vírus da Febre Hemorrágica da Crimeia-Congo/efeitos da radiação , Itália , Filogenia , Carrapatos/classificação , Vírus do Nilo Ocidental/genéticaAssuntos
Infestações por Piolhos/terapia , Refugiados/psicologia , Síndrome do Desconforto Respiratório/terapia , Recusa do Paciente ao Tratamento/psicologia , Adolescente , África Oriental/etnologia , Borrelia/patogenicidade , Infecções por Borrelia/psicologia , Infecções por Borrelia/terapia , Feminino , Humanos , Hipotensão/etiologia , Itália , Icterícia/etiologia , Leptospira/patogenicidade , Leptospirose/psicologia , Leptospirose/terapia , Infestações por Piolhos/psicologia , Síndrome do Desconforto Respiratório/etnologia , Síndrome do Desconforto Respiratório/psicologia , Recusa do Paciente ao Tratamento/etnologiaRESUMO
After the widespread use of Haemophilus influenzae type b (Hib) vaccine, H. influenzae invasive disease is now commonly due to non-encapsulated (NTHi), affecting mostly the youngest and the elderly. The objective of this study was to investigate H. influenzae nasopharyngeal carriage rate in adults with co-morbidities and possible associated risk factors. METHODS: Patients aged >50 years with co-morbidities attending medical centres were examined. A nasopharyngeal swab was analysed for H. influenzae presence by cultural and molecular methods (RT-PCR). Univariable and multivariable analysis of risk factors for H. influenzae carriage were performed. Serotype of isolates was determined by PCR capsular genotyping. Minimum inhibitory concentration (MIC) was determined by MIC gradient test and ß-lactamase production was detected by the nitrocephin test. Genotyping was performed by Multilocus sequence typing (MLST). Phylogenetic relationships among carriage and invasive NTHi strains were assessed. RESULTS: Among 248 enrolled patients (median age: 73 years), the carriage rate was 5.6% and 10.5% by cultural method or RT-PCR, respectively. Colonization with H. influenzae was significantly associated with the presence of acute respiratory symptoms (adjusted OR = 12.16, 95% CI: 3.05-48.58, p < 0.001). All colonizing isolates were NTHi. Three isolates (3/14, 21.4%) were resistant to ampicillin and beta-lactamase positive. MLST revealed a high degree of genetic diversity, with 11 different STs from 14 isolates. Eight out of the 11 (72.7%) STs were shared among carriage and invasive isolates. CONCLUSIONS: Adults ≥50 years old with co-morbidities are occasionally colonized by H. influenzae, even if the presence of co-morbidities is not a risk factor for colonization. The presence of acute respiratory symptoms is the only factor associated with H. influenzae colonization. Colonizing H. influenzae are all NTHi. Colonizing H. influenzae often belong to the same STs of invasive disease isolates.
Assuntos
Infecções por Haemophilus , Haemophilus influenzae , Adulto , Idoso , Infecções por Haemophilus/epidemiologia , Haemophilus influenzae/genética , Humanos , Lactente , Pessoa de Meia-Idade , Morbidade , Tipagem de Sequências Multilocus , Nasofaringe , FilogeniaRESUMO
BACKGROUND: The spread of carbapenems resistance is a public health concern. The main group of carbapenemases encoding the ß-lactamases activity (bla genes) is the Metallo-ß-lactamases (MBLs). METHODS: The presence of carbapenemase blaOXA-23-like, blaOXA-40-like, blaOXA-51-like, blaOXA-58-like, and blaNDM-1 genes was screened by real time PCR in 26 Pediculus humanus insects identified from second-hand clothes in a local market in Central Italy. Bacteria diversity was also characterized through shotgun metagenomic amplification for a deep sequencing of the host-associated bacterial microbiomes. RESULTS: The blaOXA-23 and blaNDM-1 carbapenemases genes were found and metagenomic analysis showed a great presence of Acinetobacter species. CONCLUSIONS: These results suggest a new potential transmission path for carbapenemase gene spread through bacteria ingested by insects infesting humans.
Assuntos
Proteínas de Bactérias/genética , Vestuário , Pediculus/genética , beta-Lactamases/genética , Animais , Humanos , Itália , Pediculus/enzimologiaRESUMO
The RNA purification is the gold standard for the detection of SARS-CoV-2 in swab samples, but it is dependent on the availability of chemical reagents. In this study, we evaluated the heat treatment method without RNA extraction as a reliable option to nucleic acid purification.
Assuntos
Betacoronavirus/genética , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Betacoronavirus/isolamento & purificação , COVID-19 , Infecções por Coronavirus/virologia , Temperatura Alta , Humanos , Pandemias , Pneumonia Viral/virologia , RNA Viral/química , RNA Viral/genética , SARS-CoV-2 , Manejo de Espécimes , Proteínas Virais/genéticaRESUMO
We report here the case of Whipple's disease in a 60-year-old man with severe arthralgia and systemic disorders but without gastrointestinal manifestations. The patient had different clinical diagnoses over a period of 14 years. We identified Tropheryma whipplei by real-time PCR. Molecular typing was also performed by sequencing the 16S-23S rRNA intergenic spacer region and domain III of the 23S rRNA gene.