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1.
PLoS Negl Trop Dis ; 14(11): e0008831, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-33166294

RESUMO

A new coronavirus (SARS-CoV-2) emerged in the winter of 2019 in Wuhan, China, and rapidly spread around the world. The extent and efficiency of SARS-CoV-2 pandemic is far greater than previous coronaviruses that emerged in the 21st Century. Here, we modeled stability of SARS-CoV-2 on skin, paper currency, and clothing to determine if these surfaces may factor in the fomite transmission dynamics of SARS-CoV-2. Skin, currency, and clothing samples were exposed to SARS-CoV-2 under laboratory conditions and incubated at three different temperatures (4°C± 2°C, 22°C± 2°C, and 37°C ± 2°C). We evaluated stability at 0 hours (h), 4 h, 8 h, 24 h, 72 h, 96 h, 7 days, and 14 days post-exposure. SARS-CoV-2 was stable on skin through the duration of the experiment at 4°C (14 days). Virus remained stable on skin for at least 96 h at 22°C and for at least 8h at 37°C. There were minimal differences between the tested currency samples. The virus remained stable on the $1 U.S.A. Bank Note for at least 96 h at 4°C while we did not detect viable virus on the $20 U.S.A. Bank Note samples beyond 72 h. The virus remained stable on both Bank Notes for at least 8 h at 22°C and 4 h at 37°C. Clothing samples were similar in stability to the currency. Viable virus remained for at least 96 h at 4°C and at least 4 h at 22°C. We did not detect viable virus on clothing samples at 37°C after initial exposure. This study confirms the inverse relationship between virus stability and temperature. Furthermore, virus stability on skin demonstrates the need for continued hand hygiene practices to minimize fomite transmission both in the general population as well as in workplaces where close contact is common.


Assuntos
Betacoronavirus/fisiologia , Infecções por Coronavirus/virologia , Pneumonia Viral/virologia , Pele/virologia , Vestuário , Infecções por Coronavirus/transmissão , Microbiologia Ambiental , Humanos , Pandemias , Pneumonia Viral/transmissão , Propriedades de Superfície , Temperatura
2.
Arch Virol ; 165(12): 2847-2856, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33034764

RESUMO

Here, we investigated the fecal, oral, blood, and skin virome of 10 laboratory rabbits using a viral metagenomic method. In the oral samples, we detected a novel polyomavirus (RabPyV), and phylogenetic analysis based on the large T antigen, VP1 and VP2 regions indicated that the novel strain might have undergone a recombination event. Recombination analysis based on related genomes confirmed that RabPyV is a multiple recombinant between rodent-like and avian-like polyomaviruses. In fecal samples, three partial or complete genome sequences of viruses belonging to the families Picobirnaviridae, Parvoviridae, Microviridae and Coronaviridae were characterized, and phylogenetic trees were constructed based on the predicted amino acid sequences of viral proteins. This study increases the amount of genetic information on viruses present in laboratory rabbits.


Assuntos
Metagenoma , Polyomavirus/isolamento & purificação , Coelhos/virologia , Proteínas Virais/genética , Vírus/classificação , Animais , Animais de Laboratório/virologia , Sangue/virologia , Fezes/virologia , Genoma Viral , Boca/virologia , Filogenia , Pele/virologia , Vírus/isolamento & purificação , Sequenciamento Completo do Genoma
3.
Gac Med Mex ; 156(4): 354-357, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32831324

RESUMO

Introduction: Reports of dermatological manifestations in patients with COVID-19 suggest a possible cutaneous tropism of SARS-CoV-2; however, the capacity of this virus to infect the skin is unknown. Objective: To determine the susceptibility of the skin to SARS-CoV-2 infection based on the expression of viral entry factors ACE2 and TMPRSS2 in this organ. Method: A comprehensive analysis of human tissue gene expression databases was carried out looking for the presence of the ACE2 and TMPRSS2 genes in the skin. mRNA expression of these genes in skin-derived human cell lines was also assessed. Results: The analyses showed high co-expression of ACE2 and TMPRSS2 in the gastrointestinal tract and kidney, but not in the skin. Only the human immortalized keratinocyte HaCaT cell line expressed detectable levels of ACE2, and no cell line originating in the skin expressed TMPRSS2. Conclusions: Our results suggest that cutaneous manifestations in patients with COVID-19 cannot be directly attributed to the virus. It is possible that cutaneous blood vessels endothelial damage, as well as the effect of circulating inflammatory mediators produced in response to the virus, are the cause of skin involvement.


Assuntos
Infecções por Coronavirus/complicações , Peptidil Dipeptidase A/genética , Pneumonia Viral/complicações , Serina Endopeptidases/genética , Dermatopatias Virais/virologia , Betacoronavirus/isolamento & purificação , Linhagem Celular , Infecções por Coronavirus/genética , Regulação da Expressão Gênica , Predisposição Genética para Doença , Humanos , Pandemias , Pneumonia Viral/genética , Pele/virologia , Dermatopatias Virais/genética , Tropismo Viral/fisiologia , Internalização do Vírus
7.
Ann Allergy Asthma Immunol ; 125(5): 528-534, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32474160

RESUMO

OBJECTIVE: To explore links between biodiversity on all scales and allergic disease as a measure of immune dysregulation. DATA SOURCES: PubMed and Web of Science were searched using the keywords biodiversity, nature relatedness, allergic disease, microbiome, noncommunicable diseases, coronavirus disease 2019, and associated terms. STUDY SELECTIONS: Studies were selected based on relevance to human health and biodiversity. RESULTS: Contact with natural environments enriches the human microbiome, promotes regulated immune responses, and protects against allergy and both acute and chronic inflammatory disorders. These important links to ecopsychological constructs of the extinction of experience, which indicates that loss of direct, personal contact with biodiversity (wildlife and the more visible elements of the natural world), might lead to emotional apathy and irresponsible behaviors toward the environment. CONCLUSION: The immune system is a useful early barometer of environmental effects and, by means of the microbiome, is a measure of the way in which our current experiences differ from our ancestral past. Although we would benefit from further research, efforts to increase direct, personal contact with biodiversity have clear benefits for multiple aspects of physical and mental health, the skin and gut microbiome, immune function, food choices, sleep, and physical activity and promote environmental responsibility.


Assuntos
Alérgenos/imunologia , Infecções por Coronavirus/prevenção & controle , Suscetibilidade a Doenças/imunologia , Hipersensibilidade/prevenção & controle , Microbiota/imunologia , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Alérgenos/administração & dosagem , Betacoronavirus/imunologia , Betacoronavirus/patogenicidade , Biodiversidade , Infecções por Coronavirus/genética , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Ecossistema , Exposição Ambiental/análise , Extinção Biológica , Trato Gastrointestinal/imunologia , Trato Gastrointestinal/microbiologia , Trato Gastrointestinal/virologia , Interação Gene-Ambiente , Humanos , Hipersensibilidade/genética , Hipersensibilidade/imunologia , Hipersensibilidade/microbiologia , Sistema Imunitário/efeitos dos fármacos , Pneumonia Viral/genética , Pneumonia Viral/imunologia , Pneumonia Viral/virologia , Pele/imunologia , Pele/microbiologia , Pele/virologia
8.
J Am Acad Dermatol ; 83(3): 870-875, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32502585

RESUMO

BACKGROUND: During the coronavirus disease 2019 pandemic, several acral chilblain-like lesions were observed in young patients with suspected, but mostly unconfirmed, infection with severe acute respiratory syndrome coronavirus 2. The histopathologic aspect of these lesions is as yet poorly known. OBJECTIVE: To investigate the pathologic features of chilblain-like lesions. METHODS: Biopsies were obtained from 17 cases of chilblain-like lesions during the coronavirus disease 2019 pandemic in France and were studied by routine histologic examination, immunohistochemistry, and direct immunofluorescence. The patients had suspected but unconfirmed infection with severe acute respiratory syndrome coronavirus 2 (negative nasopharyngeal polymerase chain reaction and serologic test results). RESULTS: Chilblain-like lesions showed many features in common with those reported in idiopathic and autoimmune-related chilblains, including epidermal necrotic keratinocytes, dermal edema, perivascular and perieccrine sweat gland lymphocytic (predominantly CD3/CD4+) inflammation, and frequent vascular changes (endothelialitis, microthromboses, fibrin deposition, and immunoreactant deposits on vessels). CONCLUSIONS: Chilblain-like lesions show histopathologic features similar to those of idiopathic and autoimmune-related chilblains, with a high rate of vascular changes and direct immunofluorescence positivity. The role of severe acute respiratory syndrome coronavirus 2 in the development of these puzzling lesions remains to be elucidated.


Assuntos
Betacoronavirus/isolamento & purificação , Pérnio/diagnóstico , Infecções por Coronavirus/complicações , Pneumonia Viral/complicações , Dermatopatias/diagnóstico , Pele/patologia , Adolescente , Adulto , Betacoronavirus/imunologia , Biópsia , Pérnio/imunologia , Pérnio/patologia , Pérnio/virologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Diagnóstico Diferencial , Feminino , Imunofluorescência , França , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/epidemiologia , Pneumonia Viral/imunologia , Pneumonia Viral/virologia , Pele/imunologia , Pele/virologia , Dermatopatias/imunologia , Dermatopatias/patologia , Dermatopatias/virologia , Dedos do Pé , Adulto Jovem
10.
Br J Dermatol ; 183(4): 729-737, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32562567

RESUMO

BACKGROUND: Chilblains ('COVID toes') are being seen with increasing frequency in children and young adults during the COVID-19 pandemic. Detailed histopathological descriptions of COVID-19 chilblains have not been reported, and causality of SARS-CoV-2 has not yet been established. OBJECTIVES: To describe the histopathological features of COVID-19 chilblains and to explore the presence of SARS-CoV-2 in the tissue. METHODS: We examined skin biopsies from seven paediatric patients presenting with chilblains during the COVID-19 pandemic. Immunohistochemistry for SARS-CoV-2 was performed in all cases and electron microscopy in one. RESULTS: Histopathology showed variable degrees of lymphocytic vasculitis ranging from endothelial swelling and endotheliitis to fibrinoid necrosis and thrombosis. Purpura, superficial and deep perivascular lymphocytic inflammation with perieccrine accentuation, oedema, and mild vacuolar interface damage were also seen. SARS-CoV-2 immunohistochemistry was positive in endothelial cells and epithelial cells of eccrine glands. Coronavirus particles were found in the cytoplasm of endothelial cells on electron microscopy. CONCLUSIONS: Although the clinical and histopathological features were similar to other forms of chilblains, the presence of viral particles in the endothelium and the histological evidence of vascular damage support a causal relation of the lesions with SARS-CoV-2. Endothelial damage induced by the virus could be the key mechanism in the pathogenesis of COVID-19 chilblains and perhaps also in a group of patients severely affected by COVID-19 presenting with features of microangiopathic damage. What is already known about this topic? Despite the high number of cases of chilblains seen during the COVID-19 pandemic, a definite causative role for SARS-CoV-2 has not yet been proven. Different pathogenetic hypotheses have been proposed, including coagulation anomalies, interferon release and external factors. What does this study add? The demonstration of SARS-CoV-2 in endothelial cells of skin biopsies by immunohistochemistry and electron microscopy confirms that these lesions are part of the spectrum of COVID-19. Virus-induced vascular damage and secondary ischaemia could explain the pathophysiology of COVID-19 chilblains. Our findings support the hypothesis that widespread endothelial infection by SARS-CoV-2 could have a pathogenetic role in the severe forms of COVID-19. Linked Comment: Wetter. Br J Dermatol 2020; 183:611.


Assuntos
Pérnio/virologia , Infecções por Coronavirus/complicações , Endotélio Vascular/patologia , Pneumonia Viral/complicações , Dermatopatias/virologia , Vasculite/virologia , Betacoronavirus/isolamento & purificação , Betacoronavirus/patogenicidade , Biópsia , Pérnio/patologia , Criança , Infecções por Coronavirus/patologia , Infecções por Coronavirus/virologia , Células Endoteliais/patologia , Células Endoteliais/ultraestrutura , Células Endoteliais/virologia , Endotélio Vascular/virologia , Humanos , Imuno-Histoquímica , Microscopia Eletrônica , Pandemias , Pneumonia Viral/patologia , Pneumonia Viral/virologia , Pele/irrigação sanguínea , Pele/patologia , Pele/virologia , Dermatopatias/patologia , Vasculite/patologia
12.
J Dermatol Sci ; 98(2): 75-81, 2020 May.
Artigo em Inglês | MEDLINE | ID: covidwho-208374

RESUMO

BACKGROUND: Various cutaneous manifestations have been observed in patients with COVID-19 infection. However, overall similarities in the clinical presentation of these dermatological manifestations have not yet been summarized. OBJECTIVE: This review aims to provide an overview of various cutaneous manifestations in patients with COVID-19 through three case reports and a literature review. METHODS: A literature search was conducted using PubMed, OVID, and Google search engines for original and review articles. Studies written in the English language that mentioned cutaneous symptoms and COVID-19 were included. RESULTS: Eighteen articles and three additional cases reported in this paper were included in this review. Of these studies, 6 are case series and 12 are case report studies. The most common cutaneous manifestation of COVID-19 was found to be maculopapular exanthem (morbilliform), presenting in 36.1% (26/72) patients. The other cutaneous manifestations included: a papulovesicular rash (34.7%, 25/72), urticaria (9.7%, 7/72), painful acral red purple papules (15.3%, 11/72) of patients, livedo reticularis lesions (2.8%, 2/72) and petechiae (1.4%, 1/72). Majority of lesions were localized on the trunk (66.7%, 50/72), however, 19.4% (14/72) of patients experienced cutaneous manifestations in the hands and feet. Skin lesion development occurred before the onset of respiratory symptoms or COVID-19 diagnosis in 12.5% (9/72) of the patients, and lesions spontaneously healed in all patients within 10 days. Majority of the studies reported no correlation between COVID-19 severity and skin lesions. CONCLUSION: Infection with COVID-19 may result in dermatological manifestations with various clinical presentations, which may aid in the timely diagnosis of this infection.


Assuntos
Betacoronavirus/patogenicidade , Infecções por Coronavirus/virologia , Pneumonia Viral/virologia , Dermatopatias Virais/virologia , Pele/virologia , Idoso , Antivirais/uso terapêutico , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/terapia , Progressão da Doença , Diagnóstico Precoce , Feminino , Humanos , Oxigenoterapia , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/terapia , Valor Preditivo dos Testes , Pele/efeitos dos fármacos , Pele/patologia , Dermatopatias Virais/diagnóstico , Dermatopatias Virais/terapia , Fatores de Tempo , Resultado do Tratamento
13.
J Dermatol Sci ; 98(2): 75-81, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32381430

RESUMO

BACKGROUND: Various cutaneous manifestations have been observed in patients with COVID-19 infection. However, overall similarities in the clinical presentation of these dermatological manifestations have not yet been summarized. OBJECTIVE: This review aims to provide an overview of various cutaneous manifestations in patients with COVID-19 through three case reports and a literature review. METHODS: A literature search was conducted using PubMed, OVID, and Google search engines for original and review articles. Studies written in the English language that mentioned cutaneous symptoms and COVID-19 were included. RESULTS: Eighteen articles and three additional cases reported in this paper were included in this review. Of these studies, 6 are case series and 12 are case report studies. The most common cutaneous manifestation of COVID-19 was found to be maculopapular exanthem (morbilliform), presenting in 36.1% (26/72) patients. The other cutaneous manifestations included: a papulovesicular rash (34.7%, 25/72), urticaria (9.7%, 7/72), painful acral red purple papules (15.3%, 11/72) of patients, livedo reticularis lesions (2.8%, 2/72) and petechiae (1.4%, 1/72). Majority of lesions were localized on the trunk (66.7%, 50/72), however, 19.4% (14/72) of patients experienced cutaneous manifestations in the hands and feet. Skin lesion development occurred before the onset of respiratory symptoms or COVID-19 diagnosis in 12.5% (9/72) of the patients, and lesions spontaneously healed in all patients within 10 days. Majority of the studies reported no correlation between COVID-19 severity and skin lesions. CONCLUSION: Infection with COVID-19 may result in dermatological manifestations with various clinical presentations, which may aid in the timely diagnosis of this infection.


Assuntos
Betacoronavirus/patogenicidade , Infecções por Coronavirus/virologia , Pneumonia Viral/virologia , Dermatopatias Virais/virologia , Pele/virologia , Idoso , Antivirais/uso terapêutico , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/terapia , Progressão da Doença , Diagnóstico Precoce , Feminino , Humanos , Oxigenoterapia , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/terapia , Valor Preditivo dos Testes , Pele/efeitos dos fármacos , Pele/patologia , Dermatopatias Virais/diagnóstico , Dermatopatias Virais/terapia , Fatores de Tempo , Resultado do Tratamento
16.
Food Microbiol ; 90: 103486, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32336365

RESUMO

The application of Campylobacter specific bacteriophages appears as a promising food safety tool for the biocontrol of this pathogen in the poultry meat production chain. However, their isolation is a complicated challenge since their occurrence appears to be low. This work assessed the efficiency of seven protocols for recovering Campylobacter phages from chicken skin samples inoculated at phage loads from 5.0 × 101 to 5.0 × 106 PFU/g. The enrichment of chicken skin in selective Bolton broth containing target isolates was the most efficient procedure, showing a low detection limit of 5.0 × 101 PFU/g and high recovery rates of up to 560%. This method's effectiveness increased as phage concentration decreased, showing its suitability for phage isolation. When this method was applied to isolate new Campylobacter phages from retail chicken skin, a total of 280 phages were recovered achieving an isolation success rate of 257%. From the 109 samples 68 resulted phage positive (62%). Chicken skin could be, therefore, considered a rich source in Campylobacter phages. This method is a simple, reproducible and efficient approach for the successful isolation of both group II and III Campylobacter specific bacteriophages, which could be helpful for the enhancement of food safety by reducing this pathogen contamination in broiler meat.


Assuntos
Bacteriófagos/isolamento & purificação , Infecções por Campylobacter/veterinária , Campylobacter/virologia , Galinhas/virologia , Pele/virologia , Virologia/métodos , Animais , Infecções por Campylobacter/microbiologia , Galinhas/microbiologia , Microbiologia de Alimentos/métodos , Inocuidade dos Alimentos/métodos , Produtos Avícolas/microbiologia , Produtos Avícolas/virologia , Pele/microbiologia
17.
PLoS One ; 15(4): e0232044, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32320441

RESUMO

Canine distemper virus (CDV) is a multi-host pathogen that can cause significant mortality in domestic, wild terrestrial and marine mammals. It is a major conservation threat in some endangered species. Infection can result in severe respiratory disease and fatal encephalitis. Diagnosis and disease monitoring in wildlife, and differentiation of CDV from rabies (a life-threatening zoonotic disease that can produce similar neurologic signs), would benefit from the availability of a portable, point-of-care (POC) diagnostic test. We therefore developed a quantitative RT-PCR assay for CDV using shelf-stable, lyophilized reagents and target-specific primers and probes for use with the handheld Biomeme two3™ qPCR thermocycler. Biomeme's extraction methodology, lyophilized reagents, and thermocycler were compared to our standard laboratory-based methods to assess sensitivity, efficiency and overall test performance. Results using a positive control plasmid for CDV showed comparable sensitivity (detection of 50 copies) and PCR efficiency between the two platforms, and CDV detection was similar between platforms when tested using a modified live CDV vaccine. Significantly higher Ct values (average Ct = 5.1 cycles) were observed using the Biomeme platform on known CDV positive animal samples. CDV detection using the Biomeme platform was similar in 25 of 26 samples from suspect CDV cases when compared to standard virology laboratory testing. One false positive was observed that was negative upon retest. The Biomeme methodology can be adapted for detection of specific targets, and this portable technology saves time by eliminating the need for local or international sample transport for laboratory-based diagnostics. However, results of our testing suggest that decreased diagnostic sensitivity (higher Ct values) relative to laboratory-based methods was observed using animal samples, so careful validation and optimization are essential. Portable qPCR platforms can empower biologists and wildlife health professionals in remote and low-resource settings, which will greatly improve our understanding of CDV disease ecology and associated conservation threats in wildlife.


Assuntos
Vírus da Cinomose Canina/genética , Cinomose/virologia , Reação em Cadeia da Polimerase em Tempo Real/instrumentação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Animais Selvagens , Áustria , Vírus da Cinomose Canina/imunologia , Congelamento , Cabelo/virologia , Nariz/virologia , Sistemas Automatizados de Assistência Junto ao Leito , RNA Viral/isolamento & purificação , Guaxinins/virologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Pele/virologia , Estados Unidos , Vacinas Atenuadas
18.
Sci Rep ; 10(1): 1612, 2020 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-32005907

RESUMO

The incidence of Merkel cell carcinoma (MCC), a rare and highly metastatic skin malignancy, has sharply increased in the last decade. Clinical biomarkers are urgently needed for MCC prognosis, treatment response monitoring, and early diagnosis of relapse. The clinical interest of circulating tumors cells (CTCs) has been validated in many solid cancers. The aim of this study was to compare CTC detection and characterization in blood samples of patients with MCC using the CellSearch System and the RosetteSep -DEPArray workflow, an innovative procedure to enrich, detect and isolate single CTCs. In preliminary experiments (using spiked MCC cell lines) both methods allowed detecting very few MCC cells. In blood samples from 19 patients with MCC at different stages, CellSearch detected MCC CTCs in 26% of patients, and the R-D workflow in 42% of patients. The detection of CTC-positive patients increased to 52% by the cumulative positivity rate of both methodologies. Moreover, Merkel cell polyomavirus DNA, involved in MCC oncogenesis, was detected in tumor biopsies, but not in all single CTCs from the same patient, reflecting the tumor heterogeneity. Our data demonstrate the possibility to detect, isolate and characterize CTCs in patients with MCC using two complementary approaches.


Assuntos
Carcinoma de Célula de Merkel/patologia , Carcinoma de Célula de Merkel/virologia , Poliomavírus das Células de Merkel/patogenicidade , Células Neoplásicas Circulantes/patologia , Neoplasias Cutâneas/patologia , Neoplasias Cutâneas/virologia , Idoso , Biomarcadores/sangue , Carcinoma de Célula de Merkel/sangue , Contagem de Células , Linhagem Celular , Linhagem Celular Tumoral , Feminino , Humanos , Masculino , Recidiva Local de Neoplasia/sangue , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/virologia , Prognóstico , Pele/patologia , Pele/virologia
19.
BMC Vet Res ; 16(1): 66, 2020 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-32085763

RESUMO

BACKGROUND: Lumpy skin disease (LSD) is an infectious viral disease of cattle caused by a Capripoxvirus. LSD has substantial economic implications, with infection resulting in permanent damage to the skin of affected animals which lowers their commercial value. In Uganda, LSD is endemic and cases of the disease are frequently reported to government authorities. This study was undertaken to molecularly characterize lumpy skin disease virus (LSDV) strains that have been circulating in Uganda between 2017 and 2018. Secondly, the study aimed to determine the phylogenetic relatedness of Ugandan LSDV sequences with published sequences, available in GenBank. RESULTS: A total of 7 blood samples and 16 skin nodule biopsies were screened for LSDV using PCR to confirm presence of LSDV nucleic acids. PCR positive samples were then characterised by amplifying the GPCR gene. These amplified genes were sequenced and phylogenetic trees were constructed. Out of the 23 samples analysed, 15 were positive for LSDV by PCR (65.2%). The LSDV GPCR sequences analysed contained the unique signatures of LSDV (A11, T12, T34, S99, and P199) which further confirmed their identity. Sequence comparison with vaccine strains revealed a 12 bp deletion unique to Ugandan outbreak strains. Phylogenetic analysis indicated that the LSDV sequences from this study clustered closely with sequences from neighboring East African countries and with LSDV strains from recent outbreaks in Europe. It was noted that the sequence diversity amongst LSDV strains from Africa was higher than diversity from Eurasia. CONCLUSION: The LSDV strains circulating in Uganda were closely related with sequences from neighboring African countries and from Eurasia. Comparison of the GPCR gene showed that outbreak strains differed from vaccine strains. This information is necessary to understand LSDV molecular epidemiology and to contribute knowledge towards the development of control strategies by the Government of Uganda.


Assuntos
Doença Nodular Cutânea/virologia , Vírus da Doença Nodular Cutânea/genética , Vírus da Doença Nodular Cutânea/isolamento & purificação , Animais , Bovinos , Surtos de Doenças/veterinária , Doença Nodular Cutânea/sangue , Doença Nodular Cutânea/epidemiologia , Vírus da Doença Nodular Cutânea/classificação , Filogenia , Reação em Cadeia da Polimerase/veterinária , Receptores de Quimiocinas/genética , Pele/virologia , Uganda/epidemiologia
20.
Diagn Microbiol Infect Dis ; 97(2): 115015, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32088016

RESUMO

We compared 2 molecular tests for detection of herpes simplex viruses 1 and 2 (HSV-1, HSV-2) and varicella-zoster virus (VZV): real-time polymerase chain reaction (RT-PCR) (Argene, BioMerieux, France) performed on an LC480 platform (Roche Applied Science, Mannheim, Germany) and isothermal amplification using a Solana HSV1 + 2/VZV assay (Quidel Corporation Worldwide Headquarters, San Diego, CA) with helicase-dependent amplification performed by a Solana® instrument. With both methods, HSV-1 was detected in 68/291 (23.4%), HSV-2 in 23/291 (7.9%), and VZV in 48/291 (16.5%) skin lesions. Both methods agreed completely only in detection of HSV-2 (kappa = 1). Concordance between Solana HSV1 + 2/VZV and RT-PCR was 98.3% (kappa = 0.95) for HSV-1 and 99.3% (kappa = 0.98) for VZV. Rapid detection of HSV-1, HSV-2, and VZV using the Solana platform is a useful method for routine diagnostics and for urgent swab samples requiring a short turnaround time.


Assuntos
Herpes Simples/diagnóstico , Técnicas de Amplificação de Ácido Nucleico , Reação em Cadeia da Polimerase em Tempo Real , Dermatopatias/diagnóstico , Infecção pelo Vírus da Varicela-Zoster/diagnóstico , DNA Viral/isolamento & purificação , Herpesviridae/classificação , Herpesviridae/isolamento & purificação , Herpesvirus Humano 3/isolamento & purificação , Humanos , Estudos Prospectivos , Estudos Retrospectivos , Pele/patologia , Pele/virologia , Dermatopatias/virologia , Temperatura
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