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1.
Sci Rep ; 14(1): 22240, 2024 09 27.
Artículo en Inglés | MEDLINE | ID: mdl-39333786

RESUMEN

This study aims to determine the presence of norovirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and bocavirus in air samples from a tertiary care hospital in Bangkok, Thailand. Air samples were collected in water using the BioSampler and concentrated using speedVac centrifugation. Based on RT-qPCR, norovirus RNA and SARS-CoV-2 RNA were detected in 13/60 (21.7%) and 3/60 (5.0%) of samples, respectively. One air sample had a weak positivity for both norovirus and SARS-CoV-2 RNAs. Detection rate of norovirus genogroup (G) II (13.3%) was higher than norovirus GI (6.7%). One air sample (1.7%) tested positive for GI and GII. The norovirus GI RNA concentration was 6.0 × 102 genome copies/m3. The norovirus GII RNA concentrations ranged from 3.4 × 101 to 5.0 × 103 genome copies/m3. Based on RT-nested PCR, norovirus GII was detected in two (3.3%) samples. All samples tested negative for GI RNA and bocavirus DNA. By phylogenetic analysis, GII.17, which is closely related to the outbreak Kawasaki308/JPN/2015 strain, was found in the RT-nested PCR-positive samples. This study highlights the potential of aerosols for norovirus and SARS-CoV-2 transmission and probably cause gastrointestinal and respiratory illnesses, respectively.


Asunto(s)
COVID-19 , Norovirus , ARN Viral , SARS-CoV-2 , Centros de Atención Terciaria , Norovirus/genética , Norovirus/aislamiento & purificación , Tailandia/epidemiología , Humanos , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , ARN Viral/genética , COVID-19/virología , COVID-19/epidemiología , COVID-19/transmisión , Microbiología del Aire , Bocavirus/genética , Bocavirus/aislamiento & purificación , Bocavirus/clasificación , Bocavirus Humano/genética , Bocavirus Humano/aislamiento & purificación , Infecciones por Caliciviridae/epidemiología , Infecciones por Caliciviridae/virología
2.
Virology ; 598: 110188, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39059190

RESUMEN

Feline bocavirus (FBoV) is a globally distributed linear, single-stranded DNA virus infect cats, currently classified into three distinct genotypes. Although FBoV can lead to systemic infections, its complete pathogenic potential remains unclear. In this study, 289 blood samples were collected from healthy cats in Harbin, revealing an overall FBoV prevalence of 12.1%. Notably, genotypes 1 and 3 of FBoV were found co-circulating among the cat population in Harbin. Additionally, recombination events were detected, particularly in the newly discovered NG/104 and DL/102 strains. Furthermore, negative selection sites were predominantly observed across the protein coding genes of FBoV. These findings suggest a co-circulation of genetically diverse FBoV strains among cats in Harbin, indicate that purifying selection is the primary driving force shaping the genomic evolution of FBoV, and also underscore the importance of comprehensive surveillance efforts to enhance our understanding of the epidemiology and evolutionary characteristics of FBoV.


Asunto(s)
Bocavirus , Enfermedades de los Gatos , Variación Genética , Genotipo , Infecciones por Parvoviridae , Filogenia , Gatos , Animales , China/epidemiología , Enfermedades de los Gatos/virología , Enfermedades de los Gatos/epidemiología , Infecciones por Parvoviridae/veterinaria , Infecciones por Parvoviridae/virología , Infecciones por Parvoviridae/epidemiología , Bocavirus/genética , Bocavirus/clasificación , Bocavirus/aislamiento & purificación , Prevalencia , Recombinación Genética , Genoma Viral , Evolución Molecular
3.
J Virol Methods ; 328: 114958, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38801834

RESUMEN

In this report, a multiplex PCR method was developed for the detection of three diarrhea-associated viruses in mink, including circovirus (MCV), bocavirus (MBoV), and enteritis virus (MEV). Three compatible sets of primers specific for each virus were designed respectively based on their conserved sequences. After optimization of the crucial factors such as primer concentration and annealing temperature in single and multiple amplification, three specific fragments were simultaneously amplified with the highest sensitivity and specificity in one PCR reaction. The fragments amplified were 259 bp (MCV),455 bp (MBoV) and 671 bp (MEV). The sensibility of this one-step multiplex PCR is about 10 times lower than that of regular singleplex PCR. There were no cross-reactions with some relevant pathogens like mink coronavirus, canine distemper virus, and aleutian mink disease virus. In our study we analyzed viral DNA in mink fecal samples by multiplex PCR assay from China, which revealed the occurrence of MCV, MBoV, and MEV as 3.1 %, 5.7 %, and 9.8 %, respectively. The testing results of multiplex PCR agreed with the singleplex PCR results with a coincidence rate of 100 %. These results indicated that the method could provide technical support for rapid detection of the three diarrhea-associated viruses, and epidemiological investigation of mink viral diarrhea.


Asunto(s)
Cartilla de ADN , Diarrea , Heces , Visón , Reacción en Cadena de la Polimerasa Multiplex , Sensibilidad y Especificidad , Animales , Visón/virología , Reacción en Cadena de la Polimerasa Multiplex/métodos , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , China , Diarrea/virología , Diarrea/veterinaria , Diarrea/diagnóstico , Cartilla de ADN/genética , Heces/virología , Circovirus/genética , Circovirus/aislamiento & purificación , Bocavirus/genética , Bocavirus/aislamiento & purificación , Virus de la Enteritis del Visón/genética , Virus de la Enteritis del Visón/aislamiento & purificación , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/veterinaria
4.
PLoS One ; 16(11): e0258680, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34731182

RESUMEN

OBJECTIVE: To evaluate the pathogenicity of a broad range of 11 possible gastroenteritis viruses, by means of statistical relationships with cases vs. controls, or Ct-values, in order to establish the most appropriate diagnostic panel for our general practitioner (GP) patients in the Netherlands (2010-2012). METHODS: Archived stool samples from 1340 cases and 1100 controls were retested using internally controlled multiplex real-time PCRs for putative pathogenic gastroenteritis viruses: adenovirus, astrovirus, bocavirus, enterovirus, norovirus GI and GII, human parechovirus, rotavirus, salivirus, sapovirus, and torovirus. RESULTS: The prevalence of any virus in symptomatic cases and asymptomatic controls was 16.6% (223/1340) and 10.2% (112/1100), respectively. Prevalence of astrovirus (adjusted odds ratio (aOR) 10.37; 95% confidence interval (CI) 1.34-80.06) and norovirus GII (aOR 3.10; CI 1.62-5.92) was significantly higher in cases versus controls. Rotavirus was encountered only in cases. We did not find torovirus and there was no statistically significant relationship with cases for salivirus (aOR 1,67; (CI) 0.43-6.54)), adenovirus non-group F (aOR 1.20; CI 0.75-1.91), bocavirus (aOR 0.85; CI 0.05-13.64), enterovirus (aOR 0.83; CI 0.50-1.37), human parechovirus (aOR 1.61; CI 0.54-4.77) and sapovirus (aOR 1.15; CI 0.67-1.98). Though adenovirus group F (aOR 6.37; CI 0.80-50.92) and norovirus GI (aOR 2.22, CI: 0.79-6.23) are known enteropathogenic viruses and were more prevalent in cases than in controls, this did not reach significance in this study. The Ct value did not discriminate between carriage and disease in PCR-positive subjects. CONCLUSIONS: In our population, diagnostic gastroenteritis tests should screen for adenovirus group F, astrovirus, noroviruses GI and GII, and rotavirus. Case-control studies as ours are lacking and should also be carried out in populations from other epidemiological backgrounds.


Asunto(s)
Infecciones por Enterovirus/diagnóstico , Heces/virología , Gastroenteritis/diagnóstico , Adenoviridae/genética , Adenoviridae/aislamiento & purificación , Adenoviridae/patogenicidad , Bocavirus/genética , Bocavirus/aislamiento & purificación , Bocavirus/patogenicidad , Preescolar , Infecciones por Enterovirus/genética , Infecciones por Enterovirus/patología , Infecciones por Enterovirus/virología , Femenino , Gastroenteritis/genética , Gastroenteritis/patología , Gastroenteritis/virología , Médicos Generales , Humanos , Lactante , Masculino , Norovirus/genética , Norovirus/aislamiento & purificación , Norovirus/patogenicidad , Pacientes , Rotavirus/genética , Rotavirus/aislamiento & purificación , Rotavirus/patogenicidad , Sapovirus/genética , Sapovirus/aislamiento & purificación , Sapovirus/patogenicidad
5.
PLoS One ; 15(12): e0243735, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33315873

RESUMEN

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.


Asunto(s)
Ruidos Respiratorios/etiología , Ruidos Respiratorios/genética , Infecciones del Sistema Respiratorio/diagnóstico , Bocavirus/genética , Bocavirus/aislamiento & purificación , Bocavirus/patogenicidad , Niño , Preescolar , Coronavirus/aislamiento & purificación , Coronavirus/patogenicidad , Humanos , Orthomyxoviridae/genética , Orthomyxoviridae/aislamiento & purificación , Orthomyxoviridae/patogenicidad , Virus de la Parainfluenza 1 Humana/genética , Virus de la Parainfluenza 1 Humana/aislamiento & purificación , Virus de la Parainfluenza 1 Humana/patogenicidad , Reacción en Cadena de la Polimerasa , Ruidos Respiratorios/fisiopatología , Sistema Respiratorio/patología , Sistema Respiratorio/virología , Infecciones del Sistema Respiratorio/genética , Infecciones del Sistema Respiratorio/virología
6.
Mol Cell Probes ; 53: 101647, 2020 10.
Artículo en Inglés | MEDLINE | ID: mdl-32777447

RESUMEN

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.


Asunto(s)
Bocavirus/aislamiento & purificación , Enfermedades de los Gatos/virología , Infecciones por Parvoviridae/diagnóstico , Proteínas no Estructurales Virales/genética , Animales , Bocavirus/clasificación , Bocavirus/genética , Gatos , Enteritis/virología , Heces/virología , Límite de Detección , Infecciones por Parvoviridae/veterinaria , Reacción en Cadena en Tiempo Real de la Polimerasa
7.
Viruses ; 12(5)2020 05 04.
Artículo en Inglés | MEDLINE | ID: mdl-32375386

RESUMEN

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.


Asunto(s)
Bocavirus/aislamiento & purificación , Enfermedades de los Gatos/virología , Diarrea/veterinaria , Parvovirinae/aislamiento & purificación , Viroma , Vómitos/veterinaria , Animales , Bocavirus/clasificación , Bocavirus/genética , Bocavirus/fisiología , Colombia Británica/epidemiología , Enfermedades de los Gatos/epidemiología , Gatos , Diarrea/epidemiología , Diarrea/virología , Brotes de Enfermedades , Heces/virología , Femenino , Genoma Viral , Masculino , Parvovirinae/clasificación , Parvovirinae/genética , Parvovirinae/fisiología , Filogenia , Vómitos/epidemiología , Vómitos/virología
8.
Arch Virol ; 165(6): 1469-1474, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32388598

RESUMEN

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.


Asunto(s)
Bocavirus/clasificación , Bocavirus/aislamiento & purificación , Infecciones por Parvoviridae/veterinaria , Infecciones por Parvoviridae/virología , Sciuridae/virología , Secuencia de Aminoácidos , Animales , China , Genoma Viral , Secuenciación de Nucleótidos de Alto Rendimiento , Metagenómica , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN
9.
Int J Legal Med ; 134(4): 1271-1274, 2020 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-32458044

RESUMEN

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.


Asunto(s)
Betacoronavirus , Infecciones por Coronavirus/patología , Pulmón/patología , Neumonía Viral/patología , Adenoviridae/genética , Adenoviridae/aislamiento & purificación , Adulto , Autopsia , Betacoronavirus/genética , Betacoronavirus/aislamiento & purificación , Bocavirus/genética , Bocavirus/aislamiento & purificación , Proteína C-Reactiva/análisis , COVID-19 , Coronavirus/genética , Coronavirus/aislamiento & purificación , Femenino , Humanos , Alphainfluenzavirus/genética , Alphainfluenzavirus/aislamiento & purificación , Betainfluenzavirus/genética , Betainfluenzavirus/aislamiento & purificación , Macrófagos/patología , Megacariocitos/patología , Metapneumovirus/genética , Metapneumovirus/aislamiento & purificación , Neutrófilos/patología , Obesidad Mórbida , Pandemias , Polipéptido alfa Relacionado con Calcitonina/sangre , Reacción en Cadena en Tiempo Real de la Polimerasa , Virus Sincitial Respiratorio Humano/genética , Virus Sincitial Respiratorio Humano/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rhinovirus/genética , Rhinovirus/aislamiento & purificación , SARS-CoV-2 , Suiza , Linfocitos T/patología
10.
Virus Genes ; 56(4): 522-526, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32306155

RESUMEN

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).


Asunto(s)
Bocavirus/aislamiento & purificación , Metagenómica , Infecciones por Parvoviridae/genética , Parvovirinae/aislamiento & purificación , Animales , Bovinos , Feto/virología , Genoma Viral/genética , Infecciones por Parvoviridae/virología , Parvovirinae/genética , Albúmina Sérica Bovina/genética
11.
Mol Cell Probes ; 51: 101544, 2020 06.
Artículo en Inglés | MEDLINE | ID: mdl-32109535

RESUMEN

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.


Asunto(s)
Bocavirus/aislamiento & purificación , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/virología , Virus de la Diarrea Epidémica Porcina/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Animales , Benzotiazoles/química , Bocavirus/genética , Coinfección , Cartilla de ADN , Diaminas/química , Virus de la Diarrea Epidémica Porcina/genética , Quinolinas/química , Sensibilidad y Especificidad , Porcinos , Temperatura de Transición
12.
Transbound Emerg Dis ; 67(1): 377-387, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31529612

RESUMEN

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.


Asunto(s)
Haplorrinos/virología , Infecciones por Parvoviridae/veterinaria , Parvovirinae/aislamiento & purificación , Primates/virología , Animales , Bocavirus/genética , Bocavirus/aislamiento & purificación , América Central/epidemiología , Heces/virología , Femenino , Humanos , Masculino , Infecciones por Parvoviridae/epidemiología , Infecciones por Parvoviridae/virología , Parvovirinae/genética , Reacción en Cadena de la Polimerasa/veterinaria , Prevalencia
13.
Virol Sin ; 35(1): 34-42, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31552610

RESUMEN

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.


Asunto(s)
Bocavirus/clasificación , Genoma Viral , Macaca mulatta/virología , Infecciones por Parvoviridae/veterinaria , Filogenia , Animales , Bocavirus/aislamiento & purificación , China/epidemiología , Heces/virología , Secuenciación de Nucleótidos de Alto Rendimiento , Infecciones por Parvoviridae/epidemiología , Proteínas Virales/genética
14.
Pol J Vet Sci ; 23(4): 481-489, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33480488

RESUMEN

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.


Asunto(s)
Bocavirus/aislamiento & purificación , Coronavirus Bovino/aislamiento & purificación , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Virus de la Parainfluenza 3 Bovina/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Animales , Bocavirus/genética , Bovinos , Coronavirus Bovino/genética , ADN Viral/aislamiento & purificación , Virus de la Parainfluenza 3 Bovina/genética , Plásmidos/genética , ARN Viral/aislamiento & purificación , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
15.
Sci Rep ; 9(1): 16367, 2019 11 08.
Artículo en Inglés | MEDLINE | ID: mdl-31705016

RESUMEN

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.


Asunto(s)
Bocavirus/aislamiento & purificación , Enfermedades de los Gatos/patología , Enteritis/veterinaria , Hemorragia Gastrointestinal/veterinaria , Infecciones por Parvoviridae/veterinaria , Recombinación Genética , Tropismo Viral , Animales , Bocavirus/clasificación , Bocavirus/genética , Enfermedades de los Gatos/epidemiología , Enfermedades de los Gatos/virología , Gatos , ADN Viral/análisis , Brotes de Enfermedades , Enteritis/epidemiología , Enteritis/virología , Hemorragia Gastrointestinal/epidemiología , Hemorragia Gastrointestinal/virología , Infecciones por Parvoviridae/epidemiología , Infecciones por Parvoviridae/virología , Filogenia , Tailandia/epidemiología
16.
Arch Virol ; 164(11): 2761-2768, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31506786

RESUMEN

A multiplex polymerase chain reaction (mPCR) assay was developed to detect and distinguish feline panleukopenia virus (FPV), feline bocavirus (FBoV) and feline astrovirus (FeAstV). Three pairs of primers were designed based on conserved regions in the genomic sequences of the three viruses and were used to specifically amplify targeted fragments of 237 bp from the VP2 gene of FPV, 465 bp from the NP1 gene of FBoV and 645 bp from the RdRp gene of FeAstV. The results showed that this mPCR assay was effective, because it could detect at least 2.25-4.04 × 104 copies of genomic DNA of the three viruses per µl, was highly specific, and had a good broad-spectrum ability to detect different genotypes of the targeted viruses. A total of 197 faecal samples that had been screened previously for FeAstV and FBoV were collected from domestic cats in northeast China and were tested for the three viruses using the newly developed mPCR assay. The total positive rate for these three viruses was 59.89% (118/197). From these samples, DNA from FPV, FBoV and FeAstV was detected in 73, 51 and 46 faecal samples, respectively. The mPCR testing results agreed with the routine PCR results with a coincidence rate of 100%. The results of this study show that this mPCR assay can simultaneously detect and differentiate FPV, FBoV and FeAstV and can be used as an easy, specific and efficient detection tool for clinical diagnosis and epidemiological investigation of these three viruses.


Asunto(s)
Bocavirus/genética , Proteínas de la Cápside/genética , Virus de la Panleucopenia Felina/genética , Mamastrovirus/genética , Reacción en Cadena de la Polimerasa Multiplex/métodos , Animales , Bocavirus/aislamiento & purificación , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/virología , Gatos , China , Cartilla de ADN/genética , Heces/virología , Virus de la Panleucopenia Felina/aislamiento & purificación , Mamastrovirus/aislamiento & purificación , Filogenia , Análisis de Secuencia de ADN
17.
Viruses ; 11(8)2019 07 31.
Artículo en Inglés | MEDLINE | ID: mdl-31370351

RESUMEN

Viruses belonging to the genus Bocaparvovirus(BoV) are a genetically diverse group of DNA viruses known to cause respiratory, enteric, and neurological diseases in animals, including humans. An intestinal sample from an alpaca (Vicugnapacos) herd with reoccurring diarrhea and respiratory disease was submitted for next-generation sequencing, revealing the presence of a BoV strain. The alpaca BoV strain (AlBoV) had a 58.58% whole genome nucleotide percent identity to a camel BoV from Dubai, belonging to a tentative ungulate BoV 8 species (UBoV8). Recombination events were lacking with other UBoV strains. The AlBoV genome was comprised of the NS1, NP1, and VP1 proteins. The NS1 protein had the highest amino acid percent identity range (57.89-67.85%) to the members of UBoV8, which was below the 85% cut-off set by the International Committee on Taxonomy of Viruses. The low NS1 amino acid identity suggests that AlBoV is a tentative new species. The whole genome, NS1, NP1, and VP1 phylogenetic trees illustrated distinct branching of AlBoV, sharing a common ancestor with UBoV8. Walker loop and Phospholipase A2 (PLA2) motifs that are vital for virus infectivity were identified in NS1 and VP1 proteins, respectively. Our study reports a novel BoV strain in an alpaca intestinal sample and highlights the need for additional BoV research.


Asunto(s)
Bocavirus/clasificación , Camélidos del Nuevo Mundo/virología , Genoma Viral , Filogenia , Animales , Bocavirus/aislamiento & purificación , Diarrea/virología , Heces/virología , Secuenciación de Nucleótidos de Alto Rendimiento , Metagenómica , Infecciones por Parvoviridae/virología , Infecciones del Sistema Respiratorio/virología , Análisis de Secuencia de ADN , Estados Unidos
18.
Arch Virol ; 164(9): 2351-2354, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31222429

RESUMEN

Porcine bocavirus (PBoV), which belongs the genus Bocaparvovirus, has been identified throughout the world. However, serological methods for detecting anti-PBoV antibodies are presently limited. In the present study, an indirect enzyme-linked immunosorbent assay (PBoV-rNP1 ELISA) based on a recombinant form of nucleoprotein 1 (NP1) of PBoV was established for investigating the seroprevalence of PBoV in 2025 serum specimens collected in north-central China from 2016 to 2018, and 42.3% of the samples tested positive for anti-PBoV IgG antibodies, indicating that the seroprevalence of PBoV is high in pig populations in China.


Asunto(s)
Anticuerpos Antivirales/sangre , Bocavirus/aislamiento & purificación , Nucleoproteínas/inmunología , Infecciones por Parvoviridae/veterinaria , Enfermedades de los Porcinos/virología , Animales , Anticuerpos Antivirales/inmunología , Bocavirus/clasificación , Bocavirus/genética , China/epidemiología , Ensayo de Inmunoadsorción Enzimática , Nucleoproteínas/genética , Infecciones por Parvoviridae/sangre , Infecciones por Parvoviridae/epidemiología , Infecciones por Parvoviridae/virología , Filogenia , Estudios Seroepidemiológicos , Porcinos , Enfermedades de los Porcinos/sangre , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/epidemiología
19.
Respir Med ; 151: 55-58, 2019 05.
Artículo en Inglés | MEDLINE | ID: mdl-31047118

RESUMEN

OBJECTIVE: This study is to investigate the status and clinical significance of respiratory viruses in bronchoalveolar lavage fluid (BALF) of children with PBB. METHODS: Sixty-eight children with PBB aged from 3 months to 5 years were enrolled and retrospectively reviewed from January 2014 to December 2017. Thirty-five children with persistent pneumonia or chronic pneumonia were matched as controls. Bronchoalveolar lavage fluid samples were collected for respiratory virus detection and bacterial culture. RESULTS: The detection rate of bacteria in BALF of children with PBB was 61.8%, which was significantly higher than that of control group (20%) (P < 0.001). The detection rate of virus in BALF of children with PBB was 23.5%, including 6 (8.8%) of rhinovirus, 4 (5.9%) of parainfluenza virus type 3, 2(2.9%) of bocavirus, 2 (2.9%) of respiratory syncytial virus 1 (1.5%) of human metapneumonia virus and 1 (1.5%) of influenza virus A. 10 cases (28.6%) of virus were detected in the control group, including 3 (8.6%) respiratory syncytial virus, 3 (8.6%) rhinovirus and 2 (5.7%) bocavirus. There was no significant difference of viral detection rate between the two groups (P = 0.577). CONCLUSION: Respiratory viruses can be detected in BALF of children with PBB, However, there is no evidence that PBB is virus-induced.


Asunto(s)
Bronquitis/microbiología , Líquido del Lavado Bronquioalveolar/microbiología , Líquido del Lavado Bronquioalveolar/virología , Bocavirus/aislamiento & purificación , Bronquitis/epidemiología , Broncomalacia/epidemiología , Estudios de Casos y Controles , Preescolar , Tos/etiología , Femenino , Humanos , Lactante , Virus de la Influenza A/aislamiento & purificación , Laringomalacia/epidemiología , Masculino , Metapneumovirus/aislamiento & purificación , Neutrófilos/metabolismo , Virus de la Parainfluenza 3 Humana/aislamiento & purificación , Ruidos Respiratorios/etiología , Virus Sincitiales Respiratorios/aislamiento & purificación , Estudios Retrospectivos , Rhinovirus/aislamiento & purificación , Traqueomalacia/epidemiología
20.
Arch Virol ; 164(2): 601-605, 2019 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-30488237

RESUMEN

In this study, we investigated the presence of canine bocaviruses (CBoVs) in fecal samples from 105 cats with diarrhea and 92 asymptomatic cats in northeast China. One fecal sample, 17CC0312, collected from an asymptomatic cat, was found to be positive for canine bocavirus 1 (CBoV1). The nearly complete genome of this virus was cloned and sequenced. The viral genome was 5,069 nucleotides (nt) in length and combined four open reading frames (ORFs) in the order 5'-NS1-ORF4-NP1-VP1/VP2-3'. The 17CC0312 virus shared more than 90.3% nucleotide sequence identity with CBoV1 reference sequences and was placed within the CBoV1 group in a phylogenetic tree based on complete genome sequences. Further phylogenetic analysis based on the deduced amino acid sequence of the VP2 gene showed that this feline CBoV1 strain belongs to CBoV1 lineage 3. These data provide the first molecular evidence of the presence of CBoV1 in a domestic cat and suggest that cats might be carriers of CBoV1.


Asunto(s)
Bocavirus/aislamiento & purificación , Enfermedades de los Gatos/virología , Genoma Viral , Infecciones por Parvoviridae/veterinaria , Animales , Secuencia de Bases , Bocavirus/clasificación , Bocavirus/genética , Gatos , China , Enfermedades de los Perros/virología , Perros , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Infecciones por Parvoviridae/virología , Filogenia
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