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1.
Arch Virol ; 164(11): 2775-2781, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31401693

RESUMO

Diagnosis and epidemiological analysis of human parvovirus B19 (hB19V) infections are essential for disease management in severely ill patients. This study aimed to evaluate the performance of an optimized NS1-VP1u nested PCR for detection and sequencing of viruses in clinical samples using 224 clinical and five reference samples. PCR sensitivity, specificity, and positive and negative predictive values were perfect (100%). While phylogenetic analysis of a 615 bp-long fragment demonstrated that the viruses in all of the samples belonged to genotype 1, this study confirmed that this optimized PCR could detect all known hB19V with high performance.


Assuntos
Proteínas do Capsídeo/genética , Eritema Infeccioso/diagnóstico , Eritema Infeccioso/epidemiologia , Parvovirus B19 Humano/genética , Proteínas não Estruturais Virais/genética , DNA Viral/genética , Eritema Infeccioso/virologia , Humanos , Filogenia , Reação em Cadeia da Polimerase/métodos
2.
Arch Virol ; 164(11): 2865-2871, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31401694

RESUMO

Phage Vp_R1 belongs to the family Podoviridae and has a C3 morphotype, with an elongated head with a diameter of 190 ± 1.1 nm and an ultrashort tail with a length of 9 ± 1.2 nm. The double-stranded DNA genome is 112.1 kb long, has a mol% G + C content of 40.3, contains 129 ORFs, and encodes four tRNAs. Phylogenetic analysis suggests that phage Vp_R1 is a novel member of the genus Kuravirus.


Assuntos
Genoma Viral/genética , Podoviridae/genética , Vibrio parahaemolyticus/virologia , Sequência de Aminoácidos , Composição de Bases/genética , DNA Viral/genética , Fases de Leitura Aberta/genética , Podoviridae/isolamento & purificação , Análise de Sequência de DNA , Proteínas Virais/genética
3.
Arch Virol ; 164(11): 2683-2690, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31428915

RESUMO

Current antiviral therapies against hepatitis B virus (HBV) infections, such as treatment with nucleos(t)ide analogs (NAs) and interferon alpha, can significantly lower HBV DNA titers, eventually to undetectable levels. However, it is still difficult to completely eliminate the stable template of HBV, the covalently closed circular DNA (cccDNA), and this contributes to viral rebound when treatment is discontinued. HBV pregenomic RNA (pgRNA), which was recently found to be present in the enveloped mature HBV viral particle in blood, is tentatively regarded, with still accumulating clinical evidence, as a novel bona fide virological marker reflecting the amount and status of cccDNA when serum HBV DNA becomes undetectable. HBV pgRNA and DNA share almost identical sequences, and it is therefore difficult to differentiate pgRNA from viral DNA using normal PCR methods. To exclude interference from viral DNA, methods for measuring pgRNA usually require a selective DNA degradation step, which is complicated and time-consuming and also compromises the accuracy of detection. In this study, we developed a simplified quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay with improved accuracy achieved by probing the polyA tail of pgRNA. Using clinical serum samples, we observed that not all patients share the same 3' sequence, suggesting slight differences between HBV strains in the way they end transcription. We then designed and evaluated a universal primer and probe set for distinguishing HBV pgRNA from HBV DNA. Our results demonstrated that a one-step qRT-PCR assay could selectively amplify HBV pgRNA from a mixture of HBV RNA and DNA, which is valuable for clinical applications.


Assuntos
Vírus da Hepatite B/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , RNA/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , DNA Viral/análise , DNA Viral/genética , Hepatite B/virologia , Humanos , RNA Viral/análise , RNA Viral/genética
4.
Arch Virol ; 164(11): 2873-2875, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31432269

RESUMO

A novel bipartite begomovirus infecting begomovirus-resistant tomato plants was detected via Illumina sequencing analysis, and its genome sequence was confirmed by Sanger sequencing. The DNA-A (2627 nt) and DNA-B (2587 nt) have a genome organization that is typical of New World bipartite begomoviruses, sharing 82.5% identity with tomato golden leaf distortion virus and 75.1% identity with sida chlorotic vein virus. Based on the current classification criteria for begomoviruses, this isolate should be considered a member of a new species, and the name "tomato interveinal chlorosis virus-2" (ToICV2) is proposed for this virus.


Assuntos
Begomovirus/classificação , Begomovirus/genética , Genoma Viral/genética , Lycopersicon esculentum/virologia , Sequência de Bases , Begomovirus/isolamento & purificação , Brasil , DNA Viral/genética , Doenças das Plantas/virologia , Análise de Sequência de DNA , Sequenciamento Completo do Genoma
5.
Arch Virol ; 164(11): 2877-2880, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31451964

RESUMO

A temperate bacteriophage, IME1320_01, was induced by mitomycin C treatment from Corynebacterium striatum. This phage possesses a double-stranded DNA genome of 40,086 bp with a G+C content of 58%. A total of 53 putative open reading frames (ORFs) were identified in its genome. BLASTn analysis revealed that IME1320_01 had the highest sequence similarity to Corynebacterium striatum strain 216, with a genome homology coverage of 44% and highest sequence identity of 95%. The termini of the phage genome was non-redundant, with a 13-nt 3'-protruding cohesive end. To the best of our knowledge, phage IME1320_01 is the first inducible phage to be identified in Corynebacterium striatum.


Assuntos
Corynebacterium/virologia , Genoma Viral/genética , Siphoviridae/genética , Ativação Viral/efeitos dos fármacos , Composição de Bases/genética , DNA Viral/genética , Mitomicina/farmacologia , Fases de Leitura Aberta/genética , Análise de Sequência de DNA , Siphoviridae/classificação , Siphoviridae/isolamento & purificação
6.
Arch Virol ; 164(11): 2715-2724, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31456086

RESUMO

Bovine coronavirus (BCoV) is a recognized cause of severe neonatal calf diarrhea, with a negative impact on animal welfare, leading to economic losses to the livestock industry. Cattle production is one of the most important economic sectors in Uruguay. The aim of this study was to determine the frequency of BCoV infections and their genetic diversity in Uruguayan calves and to describe the evolutionary history of the virus in South America. The overall detection rate of BCoV in Uruguay was 7.8% (64/824): 7.7% (60/782) in dairy cattle and 9.5% (4/42) in beef cattle. The detection rate of BCoV in samples from deceased and live calves was 10.0% (6/60) and 7.6% (58/763), respectively. Interestingly, there was a lower frequency of BCoV detection in calves born to vaccinated dams (3.3%, 8/240) than in calves born to unvaccinated dams (12.2%, 32/263) (OR: 4.02, 95%CI: 1.81-8.90; p = 0.00026). The frequency of BCoV detection was higher in colder months (11.8%, 44/373) than in warmer months (1.5%, 3/206) (OR: 9.05, 95%CI: 2.77-29.53, p = 0.000013). Uruguayan strains grouped together in two different lineages: one with Argentinean strains and the other with Brazilian strains. Both BCoV lineages were estimated to have entered Uruguay in 2013: one of them from Brazil (95%HPD interval: 2011-2014) and the other from Argentina (95%HPD interval: 2010-2014). The lineages differed by four amino acid changes, and both were divergent from the Mebus reference strain. Surveillance should be maintained to detect possible emerging strains that can clearly diverge at the antigenic level from vaccine strains.


Assuntos
Antígenos Virais/genética , Doenças dos Bovinos/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Coronavirus Bovino/isolamento & purificação , Animais , Antígenos Virais/imunologia , Argentina/epidemiologia , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/prevenção & controle , Doenças dos Bovinos/virologia , Infecções por Coronavirus/prevenção & controle , Coronavirus Bovino/genética , DNA Viral/genética , Disenteria/epidemiologia , Disenteria/veterinária , Disenteria/virologia , Variação Genética/genética , Uruguai/epidemiologia , Vacinação
7.
Rev Inst Med Trop Sao Paulo ; 61: e41, 2019 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-31432990

RESUMO

Even though there are data published on HPV epidemiology in Ecuador, the distribution of genotypes in Guayaquil, the largest city in the country, has not been previously determined in a study including including both, men and women. The present study aimed to determine the distribution of 37 HPV genotypes in genital samples from Ecuadorian men and women living in the city of Guayaquil. Genital samples included in daily diagnostic routine were analyzed by the 37 HPV GenoArray Diagnostic Kit (Hybribio® Ltd., Sheung Wan, Hong Kong). The relative frequency of detectable genotypes was determined. HPV relative frequency according to sample characteristics, including sex and age groups, was compared using c2 test. From the 800 samples (400 men and 400 women), 411 (51.38%) were positive for HPV DNA. The obtained frequency was higher among samples from men (253/400 or 63.25%) in comparison to samples from women (158/400 or 39.50%), with a p value <0.05. Samples from men showed a higher frequency of HPV genotypes 6, 16, 18 and 11, while among samples from women genotypes 39, 16, 6 and 58 were the most frequent. Considering male and female samples together, genotypes 6, 16, 39 and 11 presented the highest frequencies. HPV DNA was detected in half of the studied samples, with a higher frequency among samples from men. Genotype 39 was the most frequent among women, and ranked third when samples from men and women are analyzed together.


Assuntos
DNA Viral/genética , Papillomaviridae/genética , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/virologia , Adolescente , Adulto , Idoso , Criança , Estudos Transversais , Equador/epidemiologia , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Adulto Jovem
8.
BMC Bioinformatics ; 20(1): 389, 2019 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-31299914

RESUMO

BACKGROUND: Human papillomavirus (HPV) is a common sexually transmitted infection associated with cervical cancer that frequently occurs as a coinfection of types and subtypes. Highly similar sublineages that show over 100-fold differences in cancer risk are not distinguishable in coinfections with current typing methods. RESULTS: We describe an efficient set of computational tools, rkmh, for analyzing complex mixed infections of related viruses based on sequence data. rkmh makes extensive use of MinHash similarity measures, and includes utilities for removing host DNA and classifying reads by type, lineage, and sublineage. We show that rkmh is capable of assigning reads to their HPV type as well as HPV16 lineage and sublineages. CONCLUSIONS: Accurate read classification enables estimates of percent composition when there are multiple infecting lineages or sublineages. While we demonstrate rkmh for HPV with multiple sequencing technologies, it is also applicable to other mixtures of related sequences.


Assuntos
Coinfecção/diagnóstico , Coinfecção/virologia , Biologia Computacional/métodos , Papillomavirus Humano 16/fisiologia , Software , DNA Viral/genética , Papillomavirus Humano 16/classificação , Humanos , Infecções por Papillomavirus/virologia , Filogenia , Análise de Sequência de DNA , Fatores de Tempo
9.
J Med Microbiol ; 68(9): 1324-1329, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31355739

RESUMO

Purpose. To investigate the use of a corneal impression membrane (CIM) for the detection of herpes simplex virus type 1 (HSV-1) in suspected herpes simplex keratitis (HSK).Methodology. In the laboratory study, swabs and CIMs made from polytetrafluoroethylene were spiked with different concentrations of HSV-1. DNA was extracted and real-time PCR undertaken using two sets of primers. In the clinical study, consecutive patients presenting with suspected HSK were included. For each patient, samples were collected from corneal lesions with a swab and a CIM in random order. Clinical details were collected using a standardized clinical form and patients were categorized into probable, presumed and possible HSK.Results. There was no difference in the performance of both primer sets for all HSV-1 dilutions (P=0.83) using a CIM or between a CIM and a swab (P=0.18). In total, 110 patients were included. Overall, 73 patients (66.4 %) had probable, 20 patients (18.2 %) presumed and 17 patients (15.5 %) possible HSV-1 keratitis. The HSV-1 detection rate was significantly higher using a CIM (40/110, 36.4 %) than a swab (28/110, 25.5 %) (P=0.004). In the probable HSV keratitis group, the detection rate using a CIM was 43.8 % compared to 27.4 % for a swab (P=0.004). The cycle threshold values obtained for the conjunctival swabs were higher than those obtained for the CIMs (P<0.001).Conclusions. In suspected HSK, a CIM is a useful alternative to a swab and more likely to detect the presence of HSV-1.


Assuntos
Córnea/virologia , Herpes Simples/diagnóstico , Herpesvirus Humano 1/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Manejo de Espécimes/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Primers do DNA/genética , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
10.
Arch Virol ; 164(10): 2599-2603, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31278422

RESUMO

This work describes the characterization and genome annotation of a new lytic Enterococcus faecalis siphovirus, vB_EfaS_AL3 (referred to as AL3), isolated from wastewater samples collected in Liaoning Province, China. The genome of phage AL3 is composed of linear double-stranded DNA that is 40,789 bp in length with a G + C content of 34.84% and 61 putative protein-coding genes. Phylogenetic and comparative genomic analyses indicate that phage AL3 should be considered a novel phage.


Assuntos
Bacteriófagos/genética , Enterococcus faecalis/virologia , Genoma Viral , Filogenia , Análise de Sequência de DNA , Águas Residuárias/virologia , Bacteriólise , Composição de Bases , China , DNA/química , DNA/genética , DNA Viral/química , DNA Viral/genética , Microscopia Eletrônica de Transmissão , Anotação de Sequência Molecular , Ensaio de Placa Viral , Vírion/ultraestrutura
11.
Arch Virol ; 164(10): 2613-2616, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31321586

RESUMO

A new virus with a circular double-stranded DNA genome was discovered in green Sichuan pepper with vein clearing symptoms. Its complete genome of 8,014 bp contains three open reading frames (ORF) on the plus strand, which is typical of members of the genus Badnavirus in the family Caulimoviridae. Sequence comparisons revealed that the new virus has the highest nucleotide sequence identity with grapevine vein-clearing virus (GVCV). In particular, the identity of the two viruses in the ORF3 RT-RNase H region is 71.9%, which is below the species demarcation cutoff of 80% for badnaviruses. Phylogenetic analysis also placed the new virus with GVCV in a cluster. The virus was tentatively named "green Sichuan pepper vein clearing-associated virus" (GSPVCaV). The geographical distribution and genetic diversity of GSPVCaV were studied. Another isolate was found to be highly divergent.


Assuntos
Badnavirus/classificação , Badnavirus/isolamento & purificação , DNA Viral/genética , Filogeografia , Doenças das Plantas/virologia , Zanthoxylum/virologia , DNA/química , DNA/genética , DNA Circular/química , DNA Circular/genética , DNA Viral/química , Genoma Viral , Fases de Leitura Aberta , Análise de Sequência de DNA , Homologia de Sequência
12.
Nat Commun ; 10(1): 2753, 2019 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-31266936

RESUMO

Elimination of HIV-1 requires clearance and removal of integrated proviral DNA from infected cells and tissues. Here, sequential long-acting slow-effective release antiviral therapy (LASER ART) and CRISPR-Cas9 demonstrate viral clearance in latent infectious reservoirs in HIV-1 infected humanized mice. HIV-1 subgenomic DNA fragments, spanning the long terminal repeats and the Gag gene, are excised in vivo, resulting in elimination of integrated proviral DNA; virus is not detected in blood, lymphoid tissue, bone marrow and brain by nested and digital-droplet PCR as well as RNAscope tests. No CRISPR-Cas9 mediated off-target effects are detected. Adoptive transfer of human immunocytes from dual treated, virus-free animals to uninfected humanized mice fails to produce infectious progeny virus. In contrast, HIV-1 is readily detected following sole LASER ART or CRISPR-Cas9 treatment. These data provide proof-of-concept that permanent viral elimination is possible.


Assuntos
Fármacos Anti-HIV/administração & dosagem , Sistemas CRISPR-Cas , Infecções por HIV/terapia , HIV-1/genética , Transferência Adotiva , Animais , Terapia Combinada , DNA Viral/genética , DNA Viral/imunologia , Edição de Genes , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Infecções por HIV/virologia , HIV-1/imunologia , HIV-1/fisiologia , Humanos , Camundongos , Resultado do Tratamento , Latência Viral
13.
Nat Commun ; 10(1): 2737, 2019 06 21.
Artigo em Inglês | MEDLINE | ID: mdl-31227699

RESUMO

Little is known about the genotypic make-up of HIV-1 DNA genomes during the earliest stages of HIV-1 infection. Here, we use near-full-length, single genome next-generation sequencing to longitudinally genotype and quantify subtype C HIV-1 DNA in four women identified during acute HIV-1 infection in Durban, South Africa, through twice-weekly screening of high-risk participants. In contrast to chronically HIV-1-infected patients, we found that at the earliest phases of infection in these four participants, the majority of viral DNA genomes are intact, lack APOBEC-3G/F-associated hypermutations, have limited genome truncations, and over one year show little indication of cytotoxic T cell-driven immune selections. Viral sequence divergence during acute infection is predominantly fueled by single-base substitutions and is limited by treatment initiation during the earliest stages of disease. Our observations provide rare longitudinal insights of HIV-1 DNA sequence profiles during the first year of infection to inform future HIV cure research.


Assuntos
DNA Viral/genética , Evolução Molecular , Genoma Viral/genética , Infecções por HIV/virologia , HIV-1/genética , Doença Aguda , Adulto , Fármacos Anti-HIV/uso terapêutico , Análise Mutacional de DNA , Feminino , Seguimentos , Infecções por HIV/sangue , Infecções por HIV/tratamento farmacológico , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Estudos Longitudinais , Mutação , Estudos Prospectivos , África do Sul , Carga Viral , Adulto Jovem
14.
Arch Virol ; 164(8): 2205-2207, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31152248

RESUMO

Pathological examination of a suckling male lamb showed severe viral pneumonia with suspected bacterial superinfection. Adenovirus was detected by immunohistochemical examination of the affected lung samples. Detection of the suspected adenovirus by PCR and subsequent isolation of the virus were successful. Using next-generation sequencing, the full genome of this ovine adenovirus was sequenced and analysed. A genome sequence comparison showed that it was a novel mastadenovirus type (named "ovine adenovirus 8") that did not belong to any of the established adenovirus species. The genome is 36,206 bp long, containing 93-bp inverted terminal repeats and 29 predicted genes, including the two genus-specific genes (encoding proteins V and IX). Ovine adenovirus 8 shows the closest relationship to ovine adenovirus 6. These two viruses seem to merit the establishment of a novel ovine mastadenovirus species for them, for which we proposed the name "Ovine mastadenovirus C".


Assuntos
Adenoviridae/genética , Genoma Viral/genética , Mastadenovirus/genética , Infecções por Adenoviridae/virologia , Animais , DNA Viral/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Filogenia , Ovinos
15.
Acta Cytol ; 63(5): 385-390, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31163443

RESUMO

BACKGROUND: Uterine cervical cancer is the fourth most common female cancer in the world. In Japan, we have an apparently low rate of joining cervical cancer screening programs compared with Western countries. Furthermore, the incidence and mortality rate of cervical cancer among the younger generation has been increasing. OBJECT: The aim of this study was to assess the effectiveness of cervical cancer screening with human papillomavirus (HPV) testing and cytology in Japan. METHODS: Collaborating with Saga City government, we initiated a cervical cancer screening system consisting of HPV testing and baseline cervical cytology from April 2011 as a social experiment. A total of 17,284 participants have been screened with this new combination system. RESULTS: After HPV testing with cytology-based cervical cancer screening, the number of screenings done in women aged under 40 years has significantly increased. In addition, the number of women diagnosed with cervical intraepithelial neoplasia grade 3 has increased (25 of 14,025 vs. 146 of 23,049 under 50 years: p < 0.001). CONCLUSION: These data suggested that the introduction of HPV testing with cytology-based cervical cancer screening as an adjunct to conventional cytology resulted in better efficiency and more accurate screening among the Japanese population.


Assuntos
Neoplasia Intraepitelial Cervical/diagnóstico , DNA Viral/genética , Detecção Precoce de Câncer/métodos , Testes de DNA para Papilomavírus Humano , Teste de Papanicolaou , Papillomaviridae/genética , Neoplasias do Colo do Útero/diagnóstico , Esfregaço Vaginal , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasia Intraepitelial Cervical/patologia , Neoplasia Intraepitelial Cervical/virologia , Feminino , Humanos , Japão , Pessoa de Meia-Idade , Gradação de Tumores , Valor Preditivo dos Testes , Avaliação de Programas e Projetos de Saúde , Reprodutibilidade dos Testes , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia , Adulto Jovem
16.
Acta Cytol ; 63(5): 417-423, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31195388

RESUMO

INTRODUCTION: A causal link between infection with a high-risk strain of human papilloma virus (hrHPV) and the development of cervical squamous cell carcinoma (SCC) is well established. However, a small number of SCCs are hrHPV-negative by either HPV co-DNA testing and/or HPV-in situ hybridization (HPV-ISH) at the time of diagnosis. These apparently hrHPV-negative lesions are poorly understood, specifically whether hrHPV-positive precursor lesions exist, which would be detected through hrHPV-based screening. METHODS: A search of the pathology archives at the Johns Hopkins Hospital identified women with a diagnosis of hrHPV-negative cervical SCC on surgical specimen. All prior pathologies, including cervical cytology and surgical pathology specimens, and associated hrHPV DNA test results, p16 immunohistochemistry, and HPV-ISH were reviewed. RESULTS: A total of 25 women were identified having a surgical specimen diagnosed as SCC with either negative or equivocal HPV-ISH. Fifteen had a Pap test in the 6 months preceding a diagnosis of SCC, with cytology diagnoses as follows: high-grade squamous intraepithelial lesion n = 14/15; atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion n = 1/15. hrHPV co-testing was performed for 5 of these 15 women and was negative in 2/5 cases. Cervical biopsy was performed for 24 women. HPV-ISH testing, performed on 14 of the biopsy specimens, was negative for 11/14 patients. Of 15 specimens stained for p16, 14 were positive. CONCLUSION: A subset of patients exist in whom hrHPV is not detectable at or near the time of progression to SCC. Additional research is necessary to further describe this population and determine whether maintaining cytological screening would provide benefit.


Assuntos
Células Escamosas Atípicas do Colo do Útero/patologia , Carcinoma de Células Escamosas/patologia , Teste de Papanicolaou , Lesões Intraepiteliais Escamosas Cervicais/patologia , Neoplasias do Colo do Útero/patologia , Esfregaço Vaginal , Adulto , Idoso , Idoso de 80 Anos ou mais , Células Escamosas Atípicas do Colo do Útero/virologia , Baltimore , Biópsia , Carcinoma de Células Escamosas/cirurgia , Carcinoma de Células Escamosas/virologia , DNA Viral/genética , Bases de Dados Factuais , Feminino , Testes de DNA para Papilomavírus Humano , Humanos , Hibridização In Situ , Pessoa de Meia-Idade , Invasividade Neoplásica , Papillomaviridae/genética , Valor Preditivo dos Testes , Prognóstico , Reprodutibilidade dos Testes , Lesões Intraepiteliais Escamosas Cervicais/cirurgia , Lesões Intraepiteliais Escamosas Cervicais/virologia , Neoplasias do Colo do Útero/cirurgia , Neoplasias do Colo do Útero/virologia , Adulto Jovem
17.
Acta Cytol ; 63(5): 391-400, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31216550

RESUMO

OBJECTIVE: The exfoliative cell analyzer, LC-1000 (Sysmex Corporation, Japan), is a medical device that presents the cell proliferation index and 23 research parameters as indicators of cellular proliferative potential. The objective was to evaluate the clinical usability of qualitative assessment by LC-1000 compared with cytology, the human papillomavirus (HPV) test, and histology as gold standard. STUDY DESIGN: Women that visited 3 sites between July 2015 and March 2017 were registered. The primary endpoint in this study was the comparison between LC-1000 measurement and HPV test for sensitivity and specificity for cervical intraepithelial neoplasia 2+ (CIN2+). A tree model algorithm was newly constructed by a statistical method and its relationship with histological results was evaluated. RESULTS: The sensitivity and specificity of LC-1000 were 78.3 and 74.1%, while those of the HPV test were 94.7 and 85.4%, respectively. A tree model comprising five categories was constructed. The proportion of advanced lesions was higher with the change in the rank classification results from 1 to 5. The positive predictive values of CIN2+ in the categories 4 and 5 were high. Despite the small number of subjects, cancer was undetected in categories 1 and 2. In addition, the comparison with follow-up results in 19 women assessed as CIN1 showed that the rate of progression in the categories 3-5 was 50% (7/14); progression in the categories 1 and 2 was 0% (0/5). CONCLUSIONS: LC-1000 may be useful for cervical cancer screening as an index to qualitatively evaluate CIN and cancer based on the changes in characteristics of cells.


Assuntos
Adenocarcinoma in Situ/patologia , Carcinoma/patologia , Proliferação de Células , Neoplasia Intraepitelial Cervical/patologia , Citodiagnóstico/instrumentação , Detecção Precoce de Câncer/instrumentação , Lesões Intraepiteliais Escamosas Cervicais/patologia , Neoplasias do Colo do Útero/patologia , Adenocarcinoma in Situ/virologia , Automação Laboratorial/instrumentação , Biópsia , Carcinoma/virologia , Neoplasia Intraepitelial Cervical/virologia , DNA Viral/genética , Árvores de Decisões , Diagnóstico Diferencial , Detecção Precoce de Câncer/métodos , Desenho de Equipamento , Feminino , Testes de DNA para Papilomavírus Humano , Humanos , Japão , Teste de Papanicolaou , Papillomaviridae/genética , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Lesões Intraepiteliais Escamosas Cervicais/virologia , Neoplasias do Colo do Útero/virologia , Esfregaço Vaginal
18.
Semin Ophthalmol ; 34(4): 252-255, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31177936

RESUMO

Introduction: Infectious uveitis is a serious inflammatory condition that often causes grave ocular morbidity including permanent vision loss and damage to the structures of the eye. The most common causes of infectious uveitis include herpesviruses and Toxoplasma gondii. Traditionally, these infections have been identified and differentiated based on characteristic clinical examination findings; however, there is often overlap between these presentations and the unique cause of a given patient's infection is not always clear. Therefore, a reliable and fast method for definitively diagnosing infectious uveitis would be helpful and potentially sight-saving. Several groups have recently found experimental success with real-time multiplex polymerase chain reaction (PCR) techniques. Methods: A comprehensive review of the literature was undertaken to further understand the current state of real-time multiplex PCR and its clinical use. Search terms including "real time multiplex PCR", "infectious uveitis", and "uveitis diagnosis" were used. Appropriate English-language articles were included in this review. Results: Publications from four main groups (two from the United States, one from Japan, and one from India) citing success with real-time multiplex PCR were compared and contrasted. All four groups used the same technique to develop a highly sensitive and specific multiplex PCR analysis and found that their tests maintained high sensitivity and specificity during validation testing. These tests confirmed clinical suspicions in the majority of cases of infectious uveitis, but there were also cases of clinical misdiagnosis that were corrected based on molecular pathogen detection. These patients were then initiated on appropriate antimicrobial therapy with subsequent clinical improvement. Discussion: Real-time multiplex PCR is a highly sensitive and specific laboratory assay that allows for rapid and reliable molecular diagnosis of causative agents in infectious uveitis. This in turn facilitates swift initiation of effective therapy and prevents long-term ocular damage and vision loss.


Assuntos
Infecções Oculares , Reação em Cadeia da Polimerase Multiplex/métodos , Uveíte/diagnóstico , DNA Viral/genética , Infecções Oculares/diagnóstico , Infecções Oculares/parasitologia , Infecções Oculares/virologia , Herpesviridae/genética , Infecções por Herpesviridae/diagnóstico , Humanos , Sensibilidade e Especificidade , Toxoplasma/genética , Toxoplasmose Ocular/diagnóstico
19.
J Med Microbiol ; 68(8): 1233-1239, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31215861

RESUMO

PURPOSE: Human bocavirus (HBoV) is a DNA virus that is mostly associated with respiratory infections. However, because it has been found in stool samples, it has been suggested that it may be a causative agent for human enteric conditions. This underpins the continuous search for HBoVs, especially after the introduction of the rotavirus vaccine due to acute gastroenteritis cases related to emergent viruses, as HBoVs are more likely to be found in this post-vaccine scenario. Therefore, the aim of this study is to demonstrate the prevalence of HBoV in children aged less than 10 years with acute gastroenteritis in Brazil from November 2011 to November 2012. METHODOLOGY: Stool samples from hospitalized children ≤10 years old who presented symptoms of acute gastroenteritis were analysed for the presence of rotavirus A (RVA) by an enzyme-linked immunosorbent assay (ELISA), and for HBoV DNA by nested PCR. RESULTS: HBoV positivity was detected in 24.0 % (54/225) of samples. Two peaks of HBoV detection were observed in November 2011 and from July to September 2012. Co-infections between HBoV and rotavirus A were identified in 50.0 % (27/54) of specimens. Phylogenetic analysis identified the presence of HBoV-1 (94.8 %), HBoV-2 (2.6 %) and HBoV-3 (2.6 %) species, with only minor variations among them. CONCLUSION: Our findings provide evidence for the circulation of most HBoV genotypes (except HBoV-4) in the North Region of Brazil at a considerable rate and further investigations are necessary to improve our knowledge in the context of HBoV infections and their role in gastrointestinal diseases.


Assuntos
Gastroenterite/epidemiologia , Bocavirus Humano/genética , Epidemiologia Molecular , Infecções por Parvoviridae/epidemiologia , Doença Aguda , Brasil/epidemiologia , Proteínas do Capsídeo/genética , Criança , Pré-Escolar , Coinfecção/epidemiologia , Coinfecção/virologia , DNA Viral/genética , Fezes/virologia , Feminino , Gastroenterite/virologia , Genótipo , Bocavirus Humano/classificação , Bocavirus Humano/isolamento & purificação , Humanos , Lactente , Recém-Nascido , Masculino , Infecções por Parvoviridae/virologia , Filogenia , Reação em Cadeia da Polimerase , Prevalência , Rotavirus/imunologia , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia , Análise de Sequência de DNA
20.
J Med Microbiol ; 68(7): 1072-1080, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31162024

RESUMO

PURPOSE: Koala retrovirus (KoRV-A) is 100  % prevalent in northern Australian (Queensland and New South Wales) koala populations, where KoRV-B has been associated with Chlamydia pecorum disease and the development of lymphosarcoma. In southern populations (Victoria and South Australia), KoRV-A is less prevalent and KoRV-B has not been detected in Victoria, while the current prevalence in South Australian populations is unknown but is thought to be low. This study aimed to determine (i) the prevalence of KoRV in the two largest South Australian koala populations [Kangaroo Island (KI) and Mount Lofty Ranges (MLR)], (ii) KoRV subtype and (iii) if an association between KoRV and C. pecorum exists. METHODOLOGY: Wild koalas were sampled in KI ( n =170) between 2014 and 2017 and in MLR ( n =75) in 2016. Clinical examinations were performed, with blood collected for KoRV detection and typing by PCR. RESULTS: KoRV prevalence was 42.4  % [72/170, 95 % confidence interval (CI): 34.9-49.8  %] in KI and 65.3  % (49/75, 95 % CI: 54.6-76.1  %) in MLR. Only KoRV-A, and not KoRV-B, was detected in both populations. In MLR, there was no statistical association between KoRV and C. pecorum infection (P =0.740), or KoRV and C. pecorum disease status ( P=0.274), although KoRV-infected koalas were more likely to present with overt C. pecorum disease than subclinical infection (odds ratio: 3.15, 95 % CI: 0.91-5.39). CONCLUSION: KoRV-A is a prevalent pathogen in wild South Australian koala populations. Future studies should continue to investigate KoRV and C. pecorum associations, as the relationship is likely to be complex and to differ between the northern and southern populations.


Assuntos
Phascolarctidae/virologia , Infecções por Retroviridae/veterinária , Retroviridae/genética , Envelhecimento , Animais , Chlamydia/classificação , Chlamydia/isolamento & purificação , Infecções por Chlamydia/complicações , Infecções por Chlamydia/veterinária , DNA Viral/genética , Feminino , Genótipo , Masculino , Razão de Chances , Prevalência , Retroviridae/isolamento & purificação , Infecções por Retroviridae/complicações , Infecções por Retroviridae/epidemiologia , Infecções por Retroviridae/virologia , Fatores de Risco , Austrália do Sul/epidemiologia
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