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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.
Arch Virol ; 165(10): 2291-2299, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32754877

RESUMO

The multimammate mouse (Mastomys natalensis; M. natalensis) serves as the main reservoir for the zoonotic arenavirus Lassa virus (LASV), and this has led to considerable investigation into the distribution of LASV and other related arenaviruses in this host species. In contrast to the situation with arenaviruses, the presence of other viruses in M. natalensis remains largely unexplored. In this study, herpesviruses and polyomaviruses were identified and partially characterized by PCR methods, sequencing, and phylogenetic analysis. In tissues sampled from M. natalensis populations in Côte d'Ivoire and Mali, six new DNA viruses (four betaherpesviruses, one gammaherpesvirus and one polyomavirus) were identified. Phylogenetic analysis based on glycoprotein B amino acid sequences showed that the herpesviruses clustered with cytomegaloviruses and rhadinoviruses of multiple rodent species. The complete circular genome of the newly identified polyomavirus was amplified by PCR. Amino acid sequence analysis of the large T antigen or VP1 showed that this virus clustered with a known polyomavirus from a house mouse (species Mus musculus polyomavirus 1). These two polyomaviruses form a clade with other rodent polyomaviruses, and the newly identified virus represents the third known polyomavirus of M. natalensis. This study represents the first identification of herpesviruses and the discovery of a novel polyomavirus in M. natalensis. In contrast to arenaviruses, we anticipate that these newly identified viruses represent a low zoonotic risk due to the normally highly restricted specificity of members of these two DNA virus families to their individual mammalian host species.


Assuntos
Genoma Viral , Infecções por Herpesviridae/epidemiologia , Herpesviridae/genética , Filogenia , Infecções por Polyomavirus/epidemiologia , Polyomavirus/genética , Doenças dos Roedores/epidemiologia , África ao Sul do Saara/epidemiologia , Animais , Antígenos Virais de Tumores/genética , Proteínas do Capsídeo/genética , Reservatórios de Doenças/virologia , Herpesviridae/classificação , Herpesviridae/isolamento & purificação , Infecções por Herpesviridae/virologia , Especificidade de Hospedeiro , Tipagem Molecular , Murinae/virologia , Polyomavirus/classificação , Polyomavirus/isolamento & purificação , Infecções por Polyomavirus/virologia , Doenças dos Roedores/virologia , Proteínas do Envelope Viral/genética
4.
BMC Infect Dis ; 20(1): 488, 2020 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-32646445

RESUMO

BACKGROUND: Washington University polyomavirus (WUPyV) is a novel human polyomavirus detected in childwith acute respiratory infection in 2007. However, the relationship between WUPyV and respiratory diseases has yet to be established for lacking of a suitable in vitro culture system. METHODS: To isolate WUPyV with human airway epithelial (HAE) cells, the positive samples were incubated in HAE, and then the nucleic acid, VP1 protein and virions were detected using real-time PCR, immunofluorescence and electron microscopy respectively. RESULTS: The result showed that WUPyV could replicate effectively in HAE cells and virions with typical polyomavirus characteristics could be observed. Additionally, the entire genome sequence of the isolated strain (BJ0771) was obtained and phylogenetic analysis indicated that BJ0771 belongs to gene cluster I. CONCLUSIONS: Our findings demonstrated clinical WUPyV strain was successfully isolated for the first time in the world and this will help unravel the etiology and pathogenic mechanisms of WUPyV in respiratory infection diseases.


Assuntos
Células Epiteliais/virologia , Infecções por Polyomavirus/diagnóstico , Infecções por Polyomavirus/virologia , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Mucosa Respiratória/patologia , Infecções Respiratórias/diagnóstico , Adolescente , Proteínas do Capsídeo/genética , Polaridade Celular , Células Cultivadas , Criança , Pré-Escolar , Células Epiteliais/metabolismo , Feminino , Humanos , Masculino , Família Multigênica , Filogenia , Reação em Cadeia da Polimerase em Tempo Real , Infecções Respiratórias/virologia , Vírion/genética , Replicação Viral , Sequenciamento Completo do Genoma
5.
Cancer Immunol Immunother ; 69(8): 1615-1626, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32314041

RESUMO

BACKGROUND: The etiological role of human papillomavirus (HPV) in oropharyngeal squamous cell carcinoma (OPSCC) is confirmed. However, the role of other oncoviruses in OPSCC is unknown. MATERIALS AND METHODS: A total of 158 consecutive OPSCC patients treated with curative intent were included. DNA extracted from tumor sections was used to detect Epstein-Barr virus (EBV), HPV, and the following polyomaviruses: John Cunningham virus (JCV), Simian virus 40 (SV40), and BK virus (BKV) with PCR. In addition, p16 expression was studied by immunohistochemistry, and EBV-encoded small RNA (EBER) transcripts were localized by in situ hybridization. The effect of viral status on overall survival (OS) and disease-free survival (DFS) was analyzed. RESULTS: A total of 94/158 samples (59.5%) were HPV-positive, 29.1% contained BKV DNA, 20.3% EBV DNA, 13.9% JCV DNA, and 0.6% SV40 DNA. EBER was expressed only in stromal lymphocytes adjacent to the tumor and correlated with HPV positivity (p = 0.026). p16 expression associated only with HPV. None of the three polyomaviruses had an impact on survival. Patients with EBER-positive but HPV-negative OPSCC had significantly poorer OS and DFS than those with HPV-positive OPSCC and slightly worse prognosis compared with the patients with EBER-negative and HPV-negative OPSCC. CONCLUSION: Polyomaviruses are detectable in OPSCC but seem to have no impact on survival, whereas HPV was the strongest viral prognostic factor. EBER expression, as a sign of latent EBV infection, may have prognostic impact among patients with HPV-negative OPSCC. EBER analysis may identify a new subgroup of OPSCCs unrelated to HPV.


Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/virologia , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Infecções por Vírus Epstein-Barr/complicações , Neoplasias Orofaríngeas/virologia , Infecções por Polyomavirus/complicações , Infecções Tumorais por Vírus/complicações , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/metabolismo , Infecções por Vírus Epstein-Barr/diagnóstico , Infecções por Vírus Epstein-Barr/virologia , Feminino , Herpesvirus Humano 4/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Orofaríngeas/diagnóstico , Neoplasias Orofaríngeas/metabolismo , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/virologia , Polyomavirus/isolamento & purificação , Infecções por Polyomavirus/diagnóstico , Infecções por Polyomavirus/virologia , Prognóstico , Infecções Tumorais por Vírus/diagnóstico , Infecções Tumorais por Vírus/virologia
6.
Nat Biomed Eng ; 4(6): 601-609, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32284553

RESUMO

In organ transplantation, infection and rejection are major causes of graft loss. They are linked by the net state of immunosuppression. To diagnose and treat these conditions earlier, and to improve long-term patient outcomes, refined strategies for the monitoring of patients after graft transplantation are needed. Here, we show that a fast and inexpensive assay based on CRISPR-Cas13 accurately detects BK polyomavirus DNA and cytomegalovirus DNA from patient-derived blood and urine samples, as well as CXCL9 messenger RNA (a marker of graft rejection) at elevated levels in urine samples from patients experiencing acute kidney transplant rejection. The assay, which we adapted for lateral-flow readout, enables-via simple visualization-the post-transplantation monitoring of common opportunistic viral infections and of graft rejection, and should facilitate point-of-care post-transplantation monitoring.


Assuntos
Sistemas CRISPR-Cas , Rejeição de Enxerto/virologia , Infecções Oportunistas/diagnóstico , Patologia Molecular/métodos , Biomarcadores/sangue , Biomarcadores/urina , Quimiocina CXCL9/sangue , Quimiocina CXCL9/urina , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Citomegalovirus/genética , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/diagnóstico , DNA Viral/sangue , DNA Viral/genética , DNA Viral/urina , Humanos , Rim , Nefropatias/virologia , Transplante de Rim/efeitos adversos , Masculino , Pessoa de Meia-Idade , Testes Imediatos , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Infecções por Polyomavirus/diagnóstico , Complicações Pós-Operatórias/diagnóstico , RNA Mensageiro , Infecções Tumorais por Vírus/diagnóstico
7.
Cancer Lett ; 468: 41-47, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31605777

RESUMO

Gliomas are tumors that originate from the glial tissue, thus involving the central nervous system with varying degrees of malignancy. The most aggressive and frequent form is glioblastoma multiforme, a disease characterized by resistance to therapies, frequent recurrences, and extremely poor median survival time. Data on overall glioma case studies demonstrate clear sex disparities regarding incidence, prognosis, drug toxicity, clinical outcome, and, recently, prediction of therapeutic response. In this study, we analyze data in the literature regarding malignant glioma, mainly glioblastoma multiforme, focusing on epidemiological and clinical evaluations. Less discussed issues, such as the role of viral infections, energy metabolism, and predictive aspects concerning the possible use of dedicated therapeutic approaches for male or female patients, will be reported together with different estimated pathogenetic mechanisms underlying astrocyte transformation and glioma chemosensitivity. In this era, where personalized/precision medicine is the most important driver for targeted cancer therapies, the lines of evidence discussed herein strongly suggest that clinical approaches to malignant glioma should consider the patient's sex. Furthermore, retrospectively revising previous clinical studies considering patient sex as a crucial variable is recommended.


Assuntos
Neoplasias Encefálicas/terapia , Glioblastoma/terapia , Disparidades nos Níveis de Saúde , Recidiva Local de Neoplasia/terapia , Medicina de Precisão/métodos , Neoplasias Encefálicas/epidemiologia , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/virologia , Citomegalovirus/isolamento & purificação , Citomegalovirus/patogenicidade , Receptores ErbB/genética , Receptores ErbB/metabolismo , Feminino , Glioblastoma/epidemiologia , Glioblastoma/genética , Glioblastoma/virologia , Humanos , Incidência , Masculino , Recidiva Local de Neoplasia/epidemiologia , Recidiva Local de Neoplasia/genética , Recidiva Local de Neoplasia/virologia , Neuroglia/patologia , Neuroglia/virologia , Papillomaviridae/isolamento & purificação , Papillomaviridae/patogenicidade , Polyomavirus/isolamento & purificação , Polyomavirus/patogenicidade , Prognóstico , Fatores de Risco , Fatores Sexuais , Transdução de Sinais/genética , Telomerase/genética , Telomerase/metabolismo
9.
Virol J ; 16(1): 137, 2019 11 14.
Artigo em Inglês | MEDLINE | ID: mdl-31727090

RESUMO

BACKGROUND: Polyomaviruses (PyVs) have a wide range of hosts, from humans to fish, and their effects on hosts vary. The differences in the infection characteristics of PyV with respect to the host are assumed to be influenced by the biochemical function of the LT-Ag protein, which is related to the cytopathic effect and tumorigenesis mechanism via interaction with the host protein. METHODS: We carried out a comparative analysis of codon usage patterns of large T-antigens (LT-Ags) of PyVs isolated from various host species and their functional domains and sequence motifs. Parity rule 2 (PR2) and neutrality analysis were applied to evaluate the effects of mutation and selection pressure on codon usage bias. To investigate evolutionary relationships among PyVs, we carried out a phylogenetic analysis, and a correspondence analysis of relative synonymous codon usage (RSCU) values was performed. RESULTS: Nucleotide composition analysis using LT-Ag gene sequences showed that the GC and GC3 values of avian PyVs were higher than those of mammalian PyVs. The effective number of codon (ENC) analysis showed host-specific ENC distribution characteristics in both the LT-Ag gene and the coding sequences of its domain regions. In the avian and fish PyVs, the codon diversity was significant, whereas the mammalian PyVs tended to exhibit conservative and host-specific evolution of codon usage bias. The results of our PR2 and neutrality analysis revealed mutation bias or highly variable GC contents by showing a narrow GC12 distribution and wide GC3 distribution in all sequences. Furthermore, the calculated RSCU values revealed differences in the codon usage preference of the LT-AG gene according to the host group. A similar tendency was observed in the two functional domains used in the analysis. CONCLUSIONS: Our study showed that specific domains or sequence motifs of various PyV LT-Ags have evolved so that each virus protein interacts with host cell targets. They have also adapted to thrive in specific host species and cell types. Functional domains of LT-Ag, which are known to interact with host proteins involved in cell proliferation and gene expression regulation, may provide important information, as they are significantly related to the host specificity of PyVs.


Assuntos
Antígenos Virais de Tumores/genética , Uso do Códon , Infecções por Polyomavirus/veterinária , Infecções por Polyomavirus/virologia , Polyomavirus/genética , Motivos de Aminoácidos , Animais , Composição de Bases , Aves , Biologia Computacional , Peixes , Humanos , Mamíferos , Filogenia , Polyomavirus/isolamento & purificação
10.
Genome Biol ; 20(1): 232, 2019 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-31690338

RESUMO

The MinHash algorithm has proven effective for rapidly estimating the resemblance of two genomes or metagenomes. However, this method cannot reliably estimate the containment of a genome within a metagenome. Here, we describe an online algorithm capable of measuring the containment of genomes and proteomes within either assembled or unassembled sequencing read sets. We describe several use cases, including contamination screening and retrospective analysis of metagenomes for novel genome discovery. Using this tool, we provide containment estimates for every NCBI RefSeq genome within every SRA metagenome and demonstrate the identification of a novel polyomavirus species from a public metagenome.


Assuntos
Contaminação por DNA , Ensaios de Triagem em Larga Escala , Metagenômica/métodos , Algoritmos , Humanos , Polyomavirus/isolamento & purificação , Proteoma
11.
Vet Microbiol ; 237: 108397, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31585638

RESUMO

Aves polyomavirus 1 (APV) causes inflammatory disease in psittacine birds, especially in young budgerigar. In this study, an APV virus (SD18 strain) was isolated from a diseased psittacine birds breeding facility. The full genome (4981 bp) of SD18 was determined and analyzed. Phylogenetic analysis of full genome sequences indicated all the APV strains form two groups. The SD18 strain showed close relationship with APV isolated from Poland, however, the other Chinese strains are located in group II, which suggested different genotypes APVs are co-circulating in China. Compared with the consensus sequence of APV full genome, the SD18 strain contains 13 nucleotide mutations, and 2 unique amino acid substitutions (R179M and Q382K) located in VP2/3 and Large T proteins. To explore the pathogenicity of the virus, the SD18 strain was used to challenge 2-week-old budgerigars. All infected birds died no later than 5 days post infection, and virus was detected in multiple organs including brain, heart, ingluvies, liver, and intestine, which indicated that SD18 is fatal and causes systemic infection in young budgerigar. In vitro studies showed that SD18 replicated efficiently in CEF cells and reached the highest viral titers at 9 days post infection. Notably, replication of SD18 stimulated IFN-ß response in CEF cells and overexpression of the VP4 or VP4Delta proteins significantly inhibited IFN-ß promoter activation, which could be the strategy of APV to escape from the host innate immunity.


Assuntos
Doenças das Aves/virologia , Melopsittacus/virologia , Infecções por Polyomavirus/veterinária , Polyomavirus/isolamento & purificação , Animais , Doenças das Aves/epidemiologia , Surtos de Doenças/veterinária , Genoma Viral , Filogenia , Polyomavirus/genética , Infecções por Polyomavirus/epidemiologia , Infecções por Polyomavirus/virologia
12.
Viruses ; 11(10)2019 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-31658738

RESUMO

As the phylogenetic organization of mammalian polyomaviruses is complex and currently incompletely resolved, we aimed at a deeper insight into their evolution by identifying polyomaviruses in host orders and families that have either rarely or not been studied. Sixteen unknown and two known polyomaviruses were identified in animals that belong to 5 orders, 16 genera, and 16 species. From 11 novel polyomaviruses, full genomes could be determined. Splice sites were predicted for large and small T antigen (LTAg, STAg) coding sequences (CDS) and examined experimentally in transfected cell culture. In addition, splice sites of seven published polyomaviruses were analyzed. Based on these data, LTAg and STAg annotations were corrected for 10/86 and 74/86 published polyomaviruses, respectively. For 25 polyomaviruses, a spliced middle T CDS was observed or predicted. Splice sites that likely indicate expression of additional, alternative T antigens, were experimentally detected for six polyomaviruses. In contrast to all other mammalian polyomaviruses, three closely related cetartiodactyl polyomaviruses display two introns within their LTAg CDS. In addition, the VP2 of Glis glis (edible dormouse) polyomavirus 1 was observed to be encoded by a spliced transcript, a unique experimental finding within the Polyomaviridae family. Co-phylogenetic analyses based on LTAg CDS revealed a measurable signal of codivergence when considering all mammalian polyomaviruses, most likely driven by relatively recent codivergence events. Lineage duplication was the only other process whose influence on polyomavirus evolution was unambiguous. Finally, our analyses suggest that an update of the taxonomy of the family is required, including the creation of novel genera of mammalian and non-mammalian polyomaviruses.


Assuntos
Antígenos Virais de Tumores/genética , Mamíferos/virologia , Polyomavirus , Animais , Evolução Biológica , Classificação , Genes Virais , Genoma Viral , Humanos , Filogenia , Polyomavirus/classificação , Polyomavirus/genética , Polyomavirus/isolamento & purificação
13.
Transfusion ; 59(12): 3689-3697, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31633816

RESUMO

BACKGROUND: Human polyomaviruses (HPyVs), like herpesviruses, cause persistent infection in a large part of the population. In immunocompromised and elderly patients, PyVs cause severe diseases such as nephropathy (BK polyomavirus [BKPyV]), progressive multifocal leukoencephalopathy (JC polyomavirus [JCPyV]), and skin cancer (Merkel cell polyomavirus [MCPyV]). Like cytomegalovirus, donor-derived PyV can cause disease in kidney transplant recipients. Possibly blood components transmit PyVs as well. To study this possibility, as a first step we determined the presence of PyV DNA in Dutch blood donations. STUDY DESIGN AND METHODS: Blood donor serum samples (n = 1016) were analyzed for the presence of DNA of 14 HPyVs using HPyV species-specific quantitative polymerase chain reaction (PCR) procedures. PCR-positive samples were subjected to confirmation by sequencing. Individual PCR findings were compared with the previously reported PyV serostatus. RESULTS: MC polyomavirus DNA was detected in 39 donors (3.8%), JCPyV and TS polyomavirus (TSPyV) DNA in five donors (both 0.5%), and HPyV9 DNA in four donors (0.4%). BKPyV, WU polyomavirus (WUPyV), HPyV6, MW polyomavirus (MWPyV), and LI polyomavirus (LIPyV) DNA was detected in one or two donors. Amplicon sequencing confirmed the expected product for BKPyV, JCPyV, WUPyV, MCPyV, HPyV6, TSPyV, MWPyV, HPyV9, and LIPyV. For JCPyV a significant association was observed between detection of viral DNA and the level of specific IgG antibodies. CONCLUSION: In 5.4% of Dutch blood donors PyV DNA was detected, including DNA from pathogenic PyVs such as JCPyV. As a next step, the infectivity of PyV in donor blood and transmission via blood components to immunocompromised recipients should be investigated.


Assuntos
Doadores de Sangue/estatística & dados numéricos , DNA Viral/análise , Polyomavirus/genética , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polyomavirus/isolamento & purificação , Polyomavirus/patogenicidade , Prevalência , Adulto Jovem
14.
Arch Virol ; 164(11): 2837-2841, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31494776

RESUMO

Since January 2019, abnormal molting has been observed frequently in approximately 40-day-old Pekin ducks in China. To investigate the possible involvement of a virus, we tested the prevalence of duck circovirus (DuCV), goose hemorrhagic polyomavirus (GHPyV), and goose parvovirus (GPV) in 11 molt cases in two provinces. GPV was detected in all cases, particularly in all samples collected from the feather area. The complete genome sequences of three GPV strains were determined and found to have 52 nucleotide changes relative to GPVs associated with short beak and dwarfism syndrome of Pekin ducks. These data will enhance our understanding of GPV diversity and outcomes of GPV infection in Pekin ducks.


Assuntos
Patos/virologia , Gansos/virologia , Muda/fisiologia , Parvovirinae/isolamento & purificação , Doenças das Aves Domésticas/virologia , Animais , China/epidemiologia , Circovirus/genética , Circovirus/isolamento & purificação , Genoma Viral/genética , Parvovirinae/genética , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Doenças das Aves Domésticas/epidemiologia
15.
J Clin Virol ; 120: 6-11, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31505316

RESUMO

BACKGROUND: While the pathogenicity of the two initially identified Human Polyomaviruses (HPyVs), BK Virus (BKPyV) and JC Virus (JCPyV) has been intensely studied, there is only limited data, on whether the occurrence of the recently discovered HPyVs correlates with high level BKPyV replication and progression towards Polyomavirus associated nephropathy (PVAN). METHODS: Therefore, we performed a comprehensive longitudinal genoprevalence analysis of 13 HPyVs using a novel multiplex assay including 400 serum and 388 urine samples obtained from 99 kidney transplant recipients (KTRs), grouped by quantitative BKPyV DNA loads and evidence of manifest BKPyV associated disease (histologically verified PVAN, high urinary decoy cell levels and concurrent decrease of renal function). RESULTS: In total, 3 different non-BKPyV/JCPyV HPyVs, Human Polyomavirus 9, Merkel Cell Polyomavirus (MCPyV) and Trichodysplasia Spinulosa associated Polyomavirus were detected in 11 blood and 21 urine samples from 21 patients. Although DNAemia of these viruses occurred more frequently during high level BKPyV DNAemia and PVAN, the increase of the detection frequency due to progression of BKPyV replication did not reach statistical significance for blood samples. The positive detection rate of MCPyV in urine, however, was significantly higher during BKPyV DNAemia in 19 KTRs of our cohort who suffered from histologically verified PVAN (p = 0.005). In one individual with PVAN, continuous long-term shedding of MCPyV in urine was observed. CONCLUSION: In our cohort the recently discovered HPyVs HPyV9, TSPyV and MCPyV emerged in blood from KTRs with variable kinetics, while detection of MCPyV DNAuria occurred more frequently during BKPyV DNAemia in patients with PVAN.


Assuntos
Vírus BK/fisiologia , Nefropatias/virologia , Transplante de Rim/efeitos adversos , Infecções por Polyomavirus/diagnóstico , Polyomavirus/fisiologia , Adulto , Idoso , Vírus BK/isolamento & purificação , Estudos de Coortes , DNA Viral/sangue , DNA Viral/urina , Feminino , Humanos , Nefropatias/sangue , Nefropatias/urina , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Polyomavirus/isolamento & purificação , Infecções por Polyomavirus/sangue , Infecções por Polyomavirus/urina , Infecções por Polyomavirus/virologia , Carga Viral , Replicação Viral , Eliminação de Partículas Virais , Adulto Jovem
16.
Med Microbiol Immunol ; 208(6): 845-854, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31375897

RESUMO

Fragmented data are available on the human polyomaviruses (HPyVs) prevalence in the gastrointestinal tract. Rearrangements in the non-coding control region (NCCR) of JCPyV and BKPyV have been extensively studied and correlated to clinical outcome; instead, little information is available for KIPyV, WUPyV and MCPyV NCCRs. To get insights into the role of HPyVs in the gastrointestinal tract, we investigated JCPyV, BKPyV, KIPyV, WUPyV and MCPyV distribution among hematological patients in concomitance with gastrointestinal symptoms. In addition, NCCRs and VP1 sequences were examined to characterize the strains circulating among the enrolled patients. DNA was extracted from 62 stool samples and qPCR was carried out to detect and quantify JCPyV, BKPyV, KIPyV, WUPyV and MCPyV genomes. Positive samples were subsequently amplified and sequenced for NCCR and VP1 regions. A phylogenetic tree was constructed aligning the obtained VP1 sequences to a set of reference sequences. qPCR revealed low viral loads for all HPyVs searched. Mono and co-infections were detected. A significant correlation was found between gastrointestinal complications and KIPyV infection. Archetype-like NCCRs were found for JCPyV and BKPyV, and a high degree of NCCRs stability was observed for KIPyV, WUPyV and MCPyV. Analysis of the VP1 sequences revealed a 99% identity with the VP1 reference sequences. The study adds important information on HPyVs prevalence and persistence in the gastrointestinal tract. Gastrointestinal signs were correlated with the presence of KIPyV, although definitive conclusions cannot be drawn. HPyVs NCCRs showed a high degree of sequence stability, suggesting that sequence rearrangements are rare in this anatomical site.


Assuntos
Fezes/virologia , Variação Genética , Neoplasias Hematológicas/complicações , Infecções por Polyomavirus/epidemiologia , Infecções por Polyomavirus/virologia , Polyomavirus/isolamento & purificação , Eliminação de Partículas Virais , Adulto , Criança , Feminino , Humanos , Masculino , Filogenia , Polyomavirus/classificação , Polyomavirus/genética , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Carga Viral
17.
J Vet Diagn Invest ; 31(5): 719-725, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31423916

RESUMO

Aves polyomavirus 1, psittacine beak and feather disease virus, and psittacid herpesvirus 1 are important pathogens of psittacine birds with the potential to cause substantial morbidity and mortality. Using publically available nucleotide sequences, we developed and validated a triplex real-time PCR (rtPCR) assay to rapidly detect these 3 viruses. The assay had high analytical sensitivity, detecting <6 copies of viral DNA per reaction, and 100% analytical specificity, showing no cross-reactivity with 59 other animal pathogens. Archived formalin-fixed, paraffin-embedded tissues from psittacine birds diagnosed at postmortem as infected with each of the viruses as well as virus-negative birds were used to validate the utility of the assay. Birds were selected for the positive cohort if they showed histologic evidence of infection (i.e., characteristic inclusion bodies in tissues); birds in the negative cohort had final diagnoses unrelated to the pathogens of interest. The triplex rtPCR assay confirmed 98% of histopathology-positive cases, and also identified subclinical infections that were not observed by histologic examination, including coinfections. Birds that tested positive only by rtPCR had significantly higher cycle threshold values compared to those with histologic evidence of infection. Positive, negative, and overall percentage agreements as well as the kappa statistic between the results of the assay and histopathology were high, demonstrating the usefulness of the assay as a tool to confirm disease diagnoses, and to improve detection of subclinical infections.


Assuntos
Doenças das Aves/diagnóstico , Infecções por Vírus de DNA/veterinária , Vírus de DNA/isolamento & purificação , Herpesviridae/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/veterinária , Psittaciformes/virologia , Alphaherpesvirinae/genética , Alphaherpesvirinae/isolamento & purificação , Animais , Doenças das Aves/virologia , Infecções por Circoviridae/diagnóstico , Infecções por Circoviridae/veterinária , Infecções por Circoviridae/virologia , Circovirus/genética , Circovirus/isolamento & purificação , Infecções por Vírus de DNA/diagnóstico , Infecções por Vírus de DNA/virologia , Vírus de DNA/genética , DNA Viral , Herpesviridae/genética , Papagaios/virologia , Polyomaviridae/genética , Polyomaviridae/isolamento & purificação , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/veterinária
18.
Virchows Arch ; 475(5): 609-616, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31264036

RESUMO

Our objective was to assess the presence of three polyomaviruses, namely SV40, JCPyV, and BKPyV, and human papillomaviruses (HPV) in adenoid cystic carcinomas (ACC) of the minor salivary glands (MiSG) in the head and neck region. The study comprised 68 MiSG ACC patients operated during 1974-2012 at the Helsinki University Hospital (Helsinki, Finland). Medical records and 68 histological samples were reviewed. Polyomaviruses were detected with quantitative PCR and the DNA-positive samples were further analyzed for the presence of viral tumor T antigen (T-ag) with immunohistochemistry. HPV genotyping was performed with a Multiplex HPV Genotyping Kit. Only JCPyV DNA was found in ACC samples, being present in 7 (10.3%) out of the 68 samples. The viral load of JCPyV was low varying between 1 to 226 copies/µg DNA. The JCPyV-positive samples originated from trachea (two samples), paranasal sinuses (one), and oral cavity (two). Additionally, JCPyV positivity was found in one lung metastasis of a tracheal tumor and one local disease failure of an oral cavity tumor. Three JCPyV DNA-positive samples showed weak nuclear staining for large T-ag. In conclusion, only JCPyV but not SV40, BKPyV, or HPV was found in ACC from the upper and lower airways. JCPyV copy numbers were low which might support its role as a "hit and run agent" in ACC carcinogenesis.


Assuntos
Carcinoma Adenoide Cístico/virologia , Papillomaviridae/isolamento & purificação , Polyomavirus/isolamento & purificação , Neoplasias das Glândulas Salivares/virologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Boca/virologia , Glândulas Salivares Menores/virologia , Carga Viral , Adulto Jovem
20.
J Virol Methods ; 273: 113687, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31271791

RESUMO

A colorimetric loop-mediated isothermal amplification (LAMP) assay was developed for the rapid and specific detection of the T gene of Aves polyomavirus 1 (APyV), a causative agent of budgerigar fledgling disease (BFD) in psittacine birds. The amplification can be completed in 40 min at 60 °C, and the results can be visually detected by the naked eye using hydroxyl naphthol blue as a colorimetric indicator. The assay specifically amplified APyV DNA but not other viral and bacterial nucleic acids. The limit of detection of the assay was 5 × 102 DNA copies/reaction, which was comparable to those of previously reported conventional polymerase chain reaction assays. In the clinical evaluation, the LAMP results showed 100% concordance with those of the previously reported PCR assays with regard to specificity, sensitivity, and percentage of overall agreement, with a kappa value of 1.0. These results indicate that the developed LAMP assay will be a valuable tool for the rapid, sensitive and specific detection of APyV from BFD-suspected psittacine bird samples even in resource-limited laboratories.


Assuntos
Doenças das Aves/diagnóstico , Colorimetria/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Papagaios/virologia , Infecções por Polyomavirus/veterinária , Polyomavirus/isolamento & purificação , Infecções Tumorais por Vírus/veterinária , Animais , Compostos Azo/química , Doenças das Aves/virologia , Primers do DNA/genética , Sensibilidade e Especificidade , Temperatura
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