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1.
PLoS One ; 16(8): e0255425, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34383794

RESUMO

Canine bocaviruses (CBoVs) have been recognized as pathogens associated with intestinal diseases. Hematogenous spreading caused by CBoV has been documented and may potentiate the virus entry across the blood-brain barrier to initiate a brain infection. This study focused attention on CBoV detection in cases of encepahlopathy and attempted to determine its viral localization. A total of 107 dog brains that histologically exhibited encephalopathy (ED) were investigated for the presence of CBoVs using polymerase chain reaction (PCR). Thirty-three histologically normal brain samples from dogs were used as a control group (CD). CBoV-2 was detected in 15 ED dogs (14.02%) but not in CD dogs (p = 0.02), while no CBoV-1 and -3 were detected. Among the CBoV-2 positive dogs, brain histological changes were characterized by nonsuppurative encephalitis, with inclusion body-like materials in some brains. In situ hybridization (ISH) and transmission electron microscopy (TEM) confirmed the presence of CBoV-2 viral particles in glial cells, supporting neurotropism of this virus. ISH signals were also detected in the intestines, lymphoid organs, and the heart, suggesting both enteral and parenteral infections of this virus. Whole genome characterization and evolutionary analysis revealed genetic diversity of CBoV-2 sequences and it was varying among the different countries where the virus was detected. This study points to a possible association of CBoV-2 with encephalopathy in dogs. It also highlights the genetic diversity and cellular tropism of this virus.


Assuntos
Bocavirus , Animais , Vírus de DNA , Doenças do Cão , Cães , Infecções por Parvoviridae , Filogenia , Análise de Sequência de DNA
2.
J Vet Diagn Invest ; 33(6): 1193-1196, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34238077

RESUMO

Bovine parvovirus 1 (BPV1) is a causative agent of respiratory, gastrointestinal, and reproductive cattle diseases. We collected 149 yak diarrhea fecal samples from 9 farms in the Qinghai-Tibet Plateau. The samples were screened for BPV1 by PCR, and 2 samples were positive for BPV1. The complete genomes of these BPV1 isolates were sequenced successfully. The sequences of these 2 variants were both 5,515 bp in length and shared 96.5-96.8% identity with 2 previously reported BPV1 genomes (GenBank DQ335247, NC_001540). Twenty-six identical amino acid mutations were found in the 2 yak variants, including 7 amino acid substitutions in receptor-binding regions of the VP2 protein, and 5 amino acid substitutions in the NS1 protein C-terminal region that functions to activate transcription. The new genome sequences contribute to better understanding of the evolution and molecular characteristics of BPV1.


Assuntos
Bocavirus , Doenças dos Bovinos , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Genômica , Filogenia , Tibet
3.
Comp Immunol Microbiol Infect Dis ; 77: 101676, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34091279

RESUMO

To understand the epidemic status of feline bocavirus (FBoV) in Anhui Province, eastern China, FBoV was successfully extracted from fecal samples of domestic cats, and five complete genomes were amplified in this study. Phylogenetic analysis showed that these five strains belong to three different FBoV genotypes. Recombination analysis showed that inter- and intra-genotype recombination events occurred. Selection pressure and codon usage bias analyses indicated that FBoV-1 and FBoV-3 continuously evolve toward adaptation, and selection pressure is the main factor for codon usage bias during evolution. This study provides the first molecular evidence of FBoV prevalence in eastern China, further enriching the available information on its genetics and evolutionary characteristics and providing a basis for further research on its evolution.


Assuntos
Bocavirus , Animais , Bocavirus/genética , Gatos , China/epidemiologia , Evolução Molecular , Fezes , Genótipo , Filogenia
4.
Viruses ; 13(2)2021 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-33672786

RESUMO

Human bocavirus 1 (HBoV1) has gained attention as a gene delivery vector with its ability to infect polarized human airway epithelia and 5.5 kb genome packaging capacity. Gorilla bocavirus 1 (GBoV1) VP3 shares 86% amino acid sequence identity with HBoV1 but has better transduction efficiency in several human cell types. Here, we report the capsid structure of GBoV1 determined to 2.76 Å resolution using cryo-electron microscopy (cryo-EM) and its interaction with mouse monoclonal antibodies (mAbs) and human sera. GBoV1 shares capsid surface morphologies with other parvoviruses, with a channel at the 5-fold symmetry axis, protrusions surrounding the 3-fold axis and a depression at the 2-fold axis. A 2/5-fold wall separates the 2-fold and 5-fold axes. Compared to HBoV1, differences are localized to the 3-fold protrusions. Consistently, native dot immunoblots and cryo-EM showed cross-reactivity and binding, respectively, by a 5-fold targeted HBoV1 mAb, 15C6. Surprisingly, recognition was observed for one out of three 3-fold targeted mAbs, 12C1, indicating some structural similarity at this region. In addition, GBoV1, tested against 40 human sera, showed the similar rates of seropositivity as HBoV1. Immunogenic reactivity against parvoviral vectors is a significant barrier to efficient gene delivery. This study is a step towards optimizing bocaparvovirus vectors with antibody escape properties.


Assuntos
Anticorpos Antivirais/imunologia , Bocavirus/ultraestrutura , Capsídeo/ultraestrutura , Gorilla gorilla/virologia , Animais , Anticorpos Monoclonais/imunologia , Bocavirus/classificação , Bocavirus/genética , Bocavirus/imunologia , Capsídeo/imunologia , Reações Cruzadas , Microscopia Crioeletrônica , Bocavirus Humano/imunologia , Humanos
5.
Arch Virol ; 166(2): 451-460, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33392822

RESUMO

To investigate the epidemic profile and genetic diversity of porcine bocavirus (PBoV), 281 clinical samples, including 236 intestinal tissue samples and 45 fecal samples were collected from diarrheic piglets on 37 different pig farms in central China, and two SYBR Green I-based quantitative PCR assays were developed to detect PBoV1/2 and PBoV3/4/5, respectively. One hundred forty-eight (52.67%) of the 281 clinical samples were positive for PBoV1/2, 117 (41.63%) were positive for PBoV3/4/5, 55 (19.57%) were positive for both PBoV1/2 and PBoV3/4/5, and 86.49% (32/37) of the pig farms were positive for PBoV. Overall, the prevalence of PBoV was 74.73% (210/281) in central China. Subsequently, nearly full-length genomic sequences of two PBoV strains (designated CH/HNZM and PBoV-TY) from two different farms were determined. Phylogenetic analysis demonstrated that the two PBoV strains obtained in this study belonged to the PBoV G2 group and had a close relationship to 10 other PBoV G2 strains but differed genetically from PBoV G1, PBoV G3, and seven other bocaviruses. CH/HNZM and PBoV-TY were closely related to the PBoV strain GD18 (KJ755666), which may be derived from the PBoV strains 0912/2012 (MH558677) and 57AT-HU (KF206160) through recombination. Compared with reference strain ZJD (HM053694)-China, more amino acid variation was found in the NS1 proteins of CH/HNZM and PBoV-TY. These data extend our understanding of the molecular epidemiology and evolution of PBoV.


Assuntos
Bocavirus/genética , Infecções por Parvoviridae/virologia , Doenças dos Suínos/virologia , Animais , China , Fezes/virologia , Variação Genética/genética , Epidemiologia Molecular/métodos , Filogenia , Prevalência , Suínos
6.
PLoS One ; 15(12): e0243735, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33315873

RESUMO

INTRODUCTION: Wheezing is a major problem in children, and respiratory viruses are often believed to be the causative agent. While molecular detection tools enable identification of respiratory viruses in wheezing children, it remains unclear if and how these viruses are associated with wheezing. The objective of this systematic review is to clarify the prevalence of different respiratory viruses in children with wheezing. METHODS: We performed an electronic in Pubmed and Global Index Medicus on 01 July 2019 and manual search. We performed search of studies that have detected common respiratory viruses in children ≤18 years with wheezing. We included only studies using polymerase chain reaction (PCR) assays. Study data were extracted and the quality of articles assessed. We conducted sensitivity, subgroup, publication bias, and heterogeneity analyses using a random effects model. RESULTS: The systematic review included 33 studies. Rhinovirus, with a prevalence of 35.6% (95% CI 24.6-47.3, I2 98.4%), and respiratory syncytial virus, at 31.0% (95% CI 19.9-43.3, I2 96.4%), were the most common viruses detected. The prevalence of other respiratory viruses was as follows: human bocavirus 8.1% (95% CI 5.3-11.3, I2 84.6%), human adenovirus 7.7% (95% CI 2.6-15.0, I2 91.0%), influenza virus6.5% (95% CI 2.2-12.6, I2 92.4%), human metapneumovirus5.8% (95% CI 3.4-8.8, I2 89.0%), enterovirus 4.3% (95% CI 0.1-12.9, I2 96.2%), human parainfluenza virus 3.8% (95% CI 1.5-6.9, I2 79.1%), and human coronavirus 2.2% (95% CI 0.6-4.4, I2 79.4%). CONCLUSIONS: Our results suggest that rhinovirus and respiratory syncytial virus may contribute to the etiology of wheezing in children. While the clinical implications of molecular detection of respiratory viruses remains an interesting question, this study helps to illuminate the potential of role respiratory viruses in pediatric wheezing. REVIEW REGISTRATION: PROSPERO, CRD42018115128.


Assuntos
Sons Respiratórios/etiologia , Sons Respiratórios/genética , Infecções Respiratórias/diagnóstico , Bocavirus/genética , Bocavirus/isolamento & purificação , Bocavirus/patogenicidade , Criança , Pré-Escolar , Coronavirus/isolamento & purificação , Coronavirus/patogenicidade , Humanos , Orthomyxoviridae/genética , Orthomyxoviridae/isolamento & purificação , Orthomyxoviridae/patogenicidade , Vírus da Parainfluenza 1 Humana/genética , Vírus da Parainfluenza 1 Humana/isolamento & purificação , Vírus da Parainfluenza 1 Humana/patogenicidade , Reação em Cadeia da Polimerase , Sons Respiratórios/fisiopatologia , Sistema Respiratório/patologia , Sistema Respiratório/virologia , Infecções Respiratórias/genética , Infecções Respiratórias/virologia
7.
Mol Cell Probes ; 53: 101647, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32777447

RESUMO

Feline bocavirus-1 (FBoV-1) was first discovered in Hong Kong in 2012, and studies have indicated that the virus may cause feline hemorrhagic enteritis. Currently, there is a lack of an effective and quantitative method for FBoV-1 detection. In this study, a TaqMan-based quantitative real-time PCR (qPCR) for FBoV-1 detection was established. Primers and probes were designed to target the conserved region of the FBoV-1 NS1 gene. The sensitivity analysis indicated that the minimum detection limit was 4.57 × 101 copies/µL. The specificity test revealed no cross-reaction with seven other common feline viruses, including the same species-FBoV-2 and FBoV-3. The sensitivity of this method was 100 times higher than that of conventional PCR (cPCR). The established method showed good repeatability, with the intra-assay and inter-assay coefficients of variation of 0.18%-1.00% and 0.27%-0.45%, respectively. Furthermore, the analysis of feline feces revealed that the detection rate by qPCR was 7.0% (9/128), whereas that by cPCR was 4.7% (6/128). In conclusion, the established qPCR assay can quantitatively detect FBoV-1 with a high sensitivity, high specificity, and good reproducibility, making it a promising technique for the clinical detection of and basic and epidemiological research on FBoV-1.


Assuntos
Bocavirus/isolamento & purificação , Doenças do Gato/virologia , Infecções por Parvoviridae/diagnóstico , Proteínas não Estruturais Virais/genética , Animais , Bocavirus/classificação , Bocavirus/genética , Gatos , Enterite/virologia , Fezes/virologia , Limite de Detecção , Infecções por Parvoviridae/veterinária , Reação em Cadeia da Polimerase em Tempo Real
8.
J. med. virol ; 92(10): 1-6, Aug. 2, 2020. tab
Artigo em Inglês | LILACS, Coleciona SUS, CONASS, SES-RS | ID: biblio-1120884

RESUMO

Respiratory viral infection can cause severe disease and hospitalization, especially among children, the elderly, and patients with comorbidities. In Brazil, the official surveillance system of severe acute respiratory infection (SARI) investigates influenza A (IAV) and B (IBV) viruses, respiratory syncytial virus (RSV), adenovirus (HAdV), and parainfluenza viruses (hPIV 1­3). In Rio Grande do Sul (RS), Brazil, many fatalities associated with SARI between 2013 and 2017 occurred among patients without underlying diseases and for whom the causative agent had not been identified using official protocols. This cross­sectional study analyzed the presence of coronaviruses (HCoV), bocavirus (HBoV), metapneumovirus (hMPV), and rhinovirus in patients who died of SARI despite not having comorbidities, and that were negative for IAV, IBV, RSV, HAdV, and hPIV. Nasopharyngeal aspirates/swabs from patients were used for nucleic acid extraction. The presence of HCoVs OC43, HKU1, NL63, and 229E; HBoV; hMPV; and rhinovirus was assessed by quantitative reverse transcription­polymerase chain reaction. Clinical data were also analyzed. Between 2013 and 2017, 16 225 cases of SARI were reported in RS; 9.8% of the patients died; 20% of all fatal cases were patients without comorbidities and for whom no pathogen was detected using standard protocols. Analysis of 271 of these cases identified HCoV in nine cases; HBoV, hMPV, and rhinovirus were detected in 3, 3, and 10 cases, respectively. Of note, patients infected with HCoV were adults. Results reinforce the importance of including coronaviruses in diagnostic panels used by official surveillance systems because besides their pandemic potential, endemic HCoVs are associated to severe disease in healthy adults.


Assuntos
Humanos , Masculino , Feminino , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Adulto Jovem , Sistema Respiratório , Coronavirus , Monitoramento Epidemiológico , Infecções , Pacientes , Rhinovirus , Vírus , Viroses , Adenoviridae , Doença , Síndrome Respiratória Aguda Grave , Influenza Humana , Bocavirus
9.
Infect Genet Evol ; 85: 104421, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32580027

RESUMO

Crab-eating (Cerdocyon thous) and Pampas foxes (Lycalopex gymnocercus) are wild canids distributed in South America. Domestic dogs (Canis lupus familiaris) and wild canids may share viral pathogens, including rabies virus (RABV), canine distemper virus (CDV), and canine parvovirus 2 (CPV-2). To characterize the virome of these wild canid species, the present work evaluated the spleen and mesenteric lymph node virome of 17 crab-eating and five Pampas foxes using high-throughput sequencing (HTS). Organ samples were pooled and sequenced using an Illumina MiSeq platform. Additional PCR analyses were performed to identify the frequencies and host origin for each virus detected by HTS. Sequences more closely related to the Paramyxoviridae, Parvoviridae and Anelloviridae families were detected, as well as circular Rep-encoding single-stranded (CRESS) DNA viruses. CDV was found only in crab-eating foxes, whereas CPV-2 was found in both canid species; both viruses were closely related to sequences reported in domestic dogs from southern Brazil. Moreover, the present work reported the detection of canine bocavirus (CBoV) strains that were genetically divergent from CBoV-1 and 2 lineages. Finally, we also characterized CRESS DNA viruses and anelloviruses with marked diversity. The results of this study contribute to the body of knowledge regarding wild canid viruses that can potentially be shared with domestic canids or other species.


Assuntos
Cães/virologia , Raposas/virologia , Viroma , Vírus/classificação , Vírus/genética , Anelloviridae/classificação , Anelloviridae/genética , Animais , Bocavirus/classificação , Bocavirus/genética , Brasil , Vírus de DNA/classificação , Vírus de DNA/genética , DNA Viral , Vírus da Cinomose Canina/classificação , Vírus da Cinomose Canina/genética , Sequenciamento de Nucleotídeos em Larga Escala , Linfonodos/virologia , Metagenômica , Paramyxoviridae/classificação , Paramyxoviridae/genética , Parvoviridae/classificação , Parvoviridae/genética , Parvovirus Canino/classificação , Parvovirus Canino/genética , Filogenia , RNA Viral , Baço/virologia , Uruguai , Viroses/veterinária , Viroses/virologia , Vírus/isolamento & purificação
10.
Viruses ; 12(5)2020 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-32375386

RESUMO

An unexplained outbreak of feline diarrhea and vomiting, negative for common enteric viral and bacterial pathogens, was subjected to viral metagenomics and PCR. We characterized from fecal samples the genome of a novel chapparvovirus we named fechavirus that was shed by 8/17 affected cats and identified three different feline bocaviruses shed by 9/17 cats. Also detected were nucleic acids from attenuated vaccine viruses, members of the normal feline virome, viruses found in only one or two cases, and viruses likely derived from ingested food products. Epidemiological investigation of disease signs, time of onset, and transfers of affected cats between three facilities support a possible role for this new chapparvovirus in a highly contagious feline diarrhea and vomiting disease.


Assuntos
Bocavirus/isolamento & purificação , Doenças do Gato/virologia , Diarreia/veterinária , Parvovirinae/isolamento & purificação , Viroma , Vômito/veterinária , Animais , Bocavirus/classificação , Bocavirus/genética , Bocavirus/fisiologia , Colúmbia Britânica/epidemiologia , Doenças do Gato/epidemiologia , Gatos , Diarreia/epidemiologia , Diarreia/virologia , Surtos de Doenças , Fezes/virologia , Feminino , Genoma Viral , Masculino , Parvovirinae/classificação , Parvovirinae/genética , Parvovirinae/fisiologia , Filogenia , Vômito/epidemiologia , Vômito/virologia
11.
Int J Legal Med ; 134(4): 1271-1274, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32458044

RESUMO

In the setting of the coronavirus disease 2019 (COVID-19) pandemic, only few data regarding lung pathology induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is available, especially without medical intervention interfering with the natural evolution of the disease. We present here the first case of forensic autopsy of a COVID-19 fatality occurring in a young woman, in the community. Diagnosis was made at necropsy and lung histology showed diffuse alveolar damage, edema, and interstitial pneumonia with a geographically heterogeneous pattern, mostly affecting the central part of the lungs. This death related to COVID-19 pathology highlights the heterogeneity and severity of central lung lesions after natural evolution of the disease.


Assuntos
Betacoronavirus , Infecções por Coronavirus/patologia , Pulmão/patologia , Pneumonia Viral/patologia , Adenoviridae/genética , Adenoviridae/isolamento & purificação , Adulto , Autopsia , Betacoronavirus/genética , Betacoronavirus/isolamento & purificação , Bocavirus/genética , Bocavirus/isolamento & purificação , Proteína C-Reativa/análise , COVID-19 , Coronavirus/genética , Coronavirus/isolamento & purificação , Feminino , Humanos , Influenzavirus A/genética , Influenzavirus A/isolamento & purificação , Influenzavirus B/genética , Influenzavirus B/isolamento & purificação , Macrófagos/patologia , Megacariócitos/patologia , Metapneumovirus/genética , Metapneumovirus/isolamento & purificação , Neutrófilos/patologia , Obesidade Mórbida , Pandemias , Pró-Calcitonina/sangue , Reação em Cadeia da Polimerase em Tempo Real , Vírus Sincicial Respiratório Humano/genética , Vírus Sincicial Respiratório Humano/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rhinovirus/genética , Rhinovirus/isolamento & purificação , SARS-CoV-2 , Suíça , Linfócitos T/patologia
12.
Arch Virol ; 165(6): 1469-1474, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32388598

RESUMO

In December 2017, a squirrel (Callosciurus phayrei) died 2 days after capture in Kunming, and its intestinal tract, heart, liver, spleen, lung, and kidney were subjected to metagenomics analysis. Reassembly and verification by reverse transcription PCR of contigs generated by next-generation sequencing yielded a 5176-nt sequence, which was designated "squirrel bocaparvovirus" (SQBOV). Phylogenetic trees based on the aa sequences of NS1, NP1, and VP1 showed that SQBOV formed an independent branch in the bocaparvovirus phylogenetic tree. The amino acid sequence identity of the NS1 of SQBOV to those of other bocaparvoviruses was below the threshold of 85% that is used to demarcate species within the genus, indicating that it should be considered a member of a new bocaparvovirus species. To our knowledge, this is the first report of a bocaparvovirus in squirrels. Our findings will enable further studies of viral diversity in rodents and of the genetic diversity and host range of bocaparvoviruses.


Assuntos
Bocavirus/classificação , Bocavirus/isolamento & purificação , Infecções por Parvoviridae/veterinária , Infecções por Parvoviridae/virologia , Sciuridae/virologia , Sequência de Aminoácidos , Animais , China , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Metagenômica , Filogenia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA
13.
Virus Genes ; 56(4): 522-526, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32306155

RESUMO

A novel parvovirus was identified as a cell culture contaminant by metagenomic analysis. Droplet digital PCR (ddPCR) was used to determine viral loads in the cell culture supernatant and further analysis, by ddPCR and DNA sequencing, demonstrated that fetal bovine serum (FBS) used during cell culture was the source of the parvovirus contamination. The FBS contained ~ 50,000 copies of the novel parvovirus DNA per ml of serum. The viral DNA was resistant to DNAse digestion. Near-full length sequence of the novel parvovirus was determined. Phylogenetic analysis demonstrated that virus belongs to the Copiparvovirus genus, being most closely related to bovine parvovirus 2 (BPV2) with 41% identity with the non-structural protein NS1 and 47% identity with the virus capsid protein of BPV2. A screen of individual and pooled bovine sera identified a closely related variant of the novel virus in a second serum pool. For classification purposes, the novel virus has been designated bovine copiparvovirus species 3 isolate JB9 (bocopivirus 3-JB9).


Assuntos
Bocavirus/isolamento & purificação , Metagenômica , Infecções por Parvoviridae/genética , Parvovirinae/isolamento & purificação , Animais , Bovinos , Feto/virologia , Genoma Viral/genética , Infecções por Parvoviridae/virologia , Parvovirinae/genética , Soroalbumina Bovina/genética
14.
Virus Genes ; 56(3): 306-315, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32020392

RESUMO

Mink bocavirus 1 (MiBoV1), a novel virus detected from the feces of domestic minks in China in 2016, may be related to gastrointestinal diseases. However, its prevalence and genetic characteristics are poorly described. In this study, we examined 192 samples collected from minks in the major mink industry province from northern China. PCR results showed that 10 samples (5.2%) were positive for MiBoV1, and 60% of MiBoV1-positive samples were co-infected with Aleutian mink disease virus or mink circovirus. MiBoV1 was detected in six serum samples. Sequence analysis demonstrated that the VP2 gene of MiBoV1 was highly conserved and had low viral diversity over the VP2 region and unique nucleotide mutations. Phylogenetic analysis of the VP2 sequence demonstrated that MiBoV1 strains formed two clades and were grouped with California sea lion bocavirus, Canine bocavirus, and Feline bocavirus. Codon usage analysis revealed that most of the preferentially used codons in MiBoV1 were A- or U-ended codons, and no evident codon usage bias was found. This study provides evidence that MiBoV1 has a low prevalence in Jilin and Hebei provinces in China. Moreover, it contributes information regarding the expansion of the limited mink bocavirus sequence and determines the codon usage bias of the VP2 gene for the first time. Epidemiological surveillance is necessary to understand the importance and evolution of MiBoV1.


Assuntos
Bocavirus/genética , Proteínas do Capsídeo/genética , Uso do Códon , Vison/virologia , Substituição de Aminoácidos , Doenças dos Animais/diagnóstico , Doenças dos Animais/virologia , Animais , Bocavirus/classificação , Códon , Mutação , Infecções por Parvoviridae/veterinária , Filogenia
16.
Mol Cell Probes ; 51: 101544, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32109535

RESUMO

The duplex real-time PCR assay based on SYBR Green І was developed for detection of porcine epidemic diarrhea virus (PEDV) and porcine bocavirus (PBoV) 3/4/5 genotypes simultaneously. Two pairs of specific primers were designed targeting the N gene sequence of PEDV and VP1 gene sequence of PBoV3/4/5. PEDV and PBoV3/4/5 could be distinguished by their different melting temperatures (Tm) in one sample. The Tm value of PEDV was 83.5 °C, and the Tm value of PBoV3/4/5 was 78.5 °C, while other swine pathogens showed no specific melting peaks. The detection limits of this assay were 10 copies/µL for both PEDV and PBoV3/4/5. A total of sixty-three intestinal tissue samples were collected from piglets suffering from diarrhea, and the viral nucleic acids detected and identified by the real-time PCR assay and conventional PCR assay. The duplex real-time PCR detection results showed that the prevalence of PEDV and PBoV3/4/5 was 85.7% and 46%, respectively, and the co-infection rate of the two viruses was 28.6%. These results indicated that this duplex real-time PCR assay was a sensitive, specific and reproducible method for differentiating PEDV and PBoV3/4/5 or their co-infection.


Assuntos
Bocavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Benzotiazóis/química , Bocavirus/genética , Coinfecção , Primers do DNA , Diaminas/química , Vírus da Diarreia Epidêmica Suína/genética , Quinolinas/química , Sensibilidade e Especificidade , Suínos , Temperatura de Transição
17.
Transbound Emerg Dis ; 67(1): 377-387, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31529612

RESUMO

Parvoviruses in the genera Bocaparvovirus (HBoV), Erythroparvovirus (B19) and Tetraparvovirus (PARV4) are the only autonomous parvoviruses known to be associated with human and non-human primates based on studies and clinical cases in humans worldwide and non-human primates in Asia and Africa. Here, the presence of these agents with pathogenic potential was assessed by PCR in blood and faeces from 55 howler monkeys, 112 white-face monkeys, 3 squirrel monkeys and 127 spider monkeys in Costa Rica and El Salvador. Overall, 3.7% (11/297) of the monkeys had HboV DNA, 0.67% (2/297) had B19 DNA, and 14.1% (42/297) had PARV4 DNA, representing the first detection of these viruses in New World Primates (NWP). Sex was significantly associated with the presence of HBoV, males having greater risk up to nine times compared with females. Captivity was associated with increased prevalence for PARV4 and when all viruses were analysed together. This study provides compelling molecular evidence of parvoviruses in NWPs and underscores the importance of future research aimed at understanding how these viruses behave in natural environments of the Neotropics and what variables may favour their presence and transmission.


Assuntos
Haplorrinos/virologia , Infecções por Parvoviridae/veterinária , Parvovirinae/isolamento & purificação , Primatas/virologia , Animais , Bocavirus/genética , Bocavirus/isolamento & purificação , América Central/epidemiologia , Fezes/virologia , Feminino , Humanos , Masculino , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Parvovirinae/genética , Reação em Cadeia da Polimerase/veterinária , Prevalência
18.
Virol Sin ; 35(1): 34-42, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31552610

RESUMO

Primate bocaparvovirus (BOV) is a possible cause of respiratory disorders and gastroenteritis in humans. However, the diversity and evolution of these viruses remain largely unknown, despite the identification of a growing number of BOVs in non-human primates (NHPs). Here, we report the identification of a novel BOV (provisionally named Macaca mulatta bocaparvovirus [MmBOV]) in the feces of wild Macaca mulatta in China by viral metagenomic analysis. Seven of 400 fecal samples from Macaca mulatta were positive for MmBOV. An almost complete genome sequence of 4,831 nucleotides was obtained, which had genomic organization and protein motifs similar to human bocaviruses (HOBVs), and shared characteristically low G/C content and weak codon usage bias. Sequence analyses of NS1, NP1, and VP1 revealed that MmBOV was most closely related to HBOV4 of Primate bocaparvovirus 2 (approximately 68.4%/70.6%, 73.3%/67.6%, and 70.4%/73.1% nucleotide/amino acid identities, respectively). Additionally, phylogenetic analysis revealed that MmBOV formed an independent peripheral branch, but clustered closely with those of the Primate bocaparvovirus species in the BOV genus (particularly HBOV4). These data strongly suggest that HBOV4 originated from NHP bocaparvoviruses around 200-300 years ago, and that NHPs may act as HBOV reservoirs. Following the International Committee of Taxonomy for Viruses guidelines, we propose MmBOV as a new species (tentatively named Primate bocaparvovirus 3) in the genus Bocaparvovirus, which is the first report of a novel species of primate BOV. Our data facilitate future research on the genetic diversity and evolution of primate bocaparvoviruses and highlight the importance of bocaparvovirus surveys in wild NHPs.


Assuntos
Bocavirus/classificação , Genoma Viral , Macaca mulatta/virologia , Infecções por Parvoviridae/veterinária , Filogenia , Animais , Bocavirus/isolamento & purificação , China/epidemiologia , Fezes/virologia , Sequenciamento de Nucleotídeos em Larga Escala , Infecções por Parvoviridae/epidemiologia , Proteínas Virais/genética
19.
Pol J Vet Sci ; 23(4): 481-489, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33480488

RESUMO

Bovine parvovirus (BPV), bovine coronavirus (BCoV) and bovine parainfluenza virus (BPIV) are common etiologies causing gastrointestinal and respiratory diseases in dairy herds. However, there are few reports on the synchronous detection of BPV, BCoV and BPIV. The present article aimed to develop a quick and accurate RT-PCR assay to synchronously detect BPV, BCoV and BPIV based on their specific probes. One pair universal primers, one pair specific primers and one specific probe was designed and synthesized. After the concentrations of primer and probe and annealing temperature were strictly optimized, the specificity, sensitivity and repeatability of the established triplex probe qRT-PCR were evaluated, respectively. The results showed the recombinant plasmids of pMD18-T-BPV, pMD18-T-BCoV and pMD18-T-BPIV were 554bp, 699bp and 704bp, respectively. The optimal annealing temperature was set at 45.0°C for triplex qRT-PCR. The triplex probe qRT-PCR can only synchronously detect BPV, BCoV and BPIV. Detection sensitivities were 2.0×102, 2.0×102 and 2.0×101 copies/µL for BPV, BCoV and BPIV, being 1000-fold greater than that in the conventional PCR. Detection of clinical samples demonstrated that triplex probe qRT-PCR had a higher sensitivity and specificity. The intra-assay and inter-assay coefficient of variation were lower than 2.0%. Clinical specimens verified that the triplex qRT-PCR had a higher sensitivity and specificity than universal PCR. In conclusion, this triplex probe qRT-PCR could detect only BPV, BCoV and BPIV. Minimum detection limits were 2.0×102 copies/µL for BPV and BCoV, and 2.0×101 copies/µL for BPIV. The sensitivity of this triplex probe qRT-PCR was 1000-fold greater than that in the conventional PCR. The newly qRT-PCR could be used to monitor or differentially diagnose virus infection.


Assuntos
Bocavirus/isolamento & purificação , Coronavirus Bovino/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/veterinária , Vírus da Parainfluenza 3 Bovina/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Animais , Bocavirus/genética , Bovinos , Coronavirus Bovino/genética , DNA Viral/isolamento & purificação , Vírus da Parainfluenza 3 Bovina/genética , Plasmídeos/genética , RNA Viral/isolamento & purificação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
20.
Sci Rep ; 9(1): 16367, 2019 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-31705016

RESUMO

Feline bocavirus-1 (FBoV-1) was identified in cats from different households with hemorrhagic enteritis during outbreaks of an unusual clinical presentation of feline panleukopenia virus (FPLV) in Thailand. Use of polymerase chain reaction revealed the presence of the FBoV-1 DNA in several tissues, suggesting hematogenous viremia, with the viral nucleic acid, detected by in situ hybridization (ISH), was localized in intestinal cells and vascular endothelium of intestinal mucosa and serosa, and in necrosis areas primarily in various lymph nodes while FPLV-immunohistochemical analysis revealed viral localization only in cryptal cells, neurons, and limited to leukocytes in the mesenteric lymph node. Full-length coding genome analysis of the Thai FBoV-1 strains isolated from moribund cats revealed three distinct strains with a high between-strain genetic diversity, while genetic recombination in one of the three FBoV-1 strains within the NS1 gene. This is the first report identifying natural genetic recombination of the FBoV-1 and describing the pathology and viral tropism of FBoV-1 infection in cats. Although the role of FBoV-1 associated with systemic infection of these cats remained undetermined, a contributory role of enteric infection of FBoV-1 is possible. Synergistic effects of dual infection with FPLV and FBoV-1 are hypothesized, suggesting more likely severe clinical presentations.


Assuntos
Bocavirus/isolamento & purificação , Doenças do Gato/patologia , Enterite/veterinária , Hemorragia Gastrointestinal/veterinária , Infecções por Parvoviridae/veterinária , Recombinação Genética , Tropismo Viral , Animais , Bocavirus/classificação , Bocavirus/genética , Doenças do Gato/epidemiologia , Doenças do Gato/virologia , Gatos , DNA Viral/análise , Surtos de Doenças , Enterite/epidemiologia , Enterite/virologia , Hemorragia Gastrointestinal/epidemiologia , Hemorragia Gastrointestinal/virologia , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Filogenia , Tailândia/epidemiologia
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