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1.
J Zoo Wildl Med ; 50(4): 769-777, 2020 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-31926506

RESUMO

Eastern box turtles (Terrapene carolina carolina) are a native North American species with a declining population trend that may be attributable to habitat fragmentation, vehicle collisions, and disease. Adenoviral infections can cause significant morbidity and mortality in captive reptile populations. Adenoviruses have been documented in box turtles, but their occurrence and impact in wild populations are unknown. A disease survey was performed at The Wildlife Center of Virginia, USA, to assess the prevalence of box turtle adenovirus (BTAdV) in wild eastern box turtles and evaluate potential associations with clinical disease. Swabs from the oral cavity, including the choanal slit, and the cloaca were collected from 106 eastern box turtles from July 2015 through June 2016. The quantitative polymerase chain reaction (qPCR) primer detected both ornate box turtle adenovirus 1 and eastern box turtle adenovirus. The resulting qPCR adenovirus prevalence was 55.7% (n = 59). Most animals (99.3%) that tested positive for BTAdV had fewer than 100 viral copies/ng DNA. This study did not find a statistically significant association between cause of admission, age, sex, outcome, and BTAdV qPCR status. However, the probability of BTAdV detection was 1.5 times higher in rehabilitation turtles compared with wild turtles (P = 0.01). Albumin was significantly lower in qPCR BTAdV-positive turtles (P = 0.007). Hypoalbuminemia is not generally associated with adenovirus infections in other species, and no obvious clinical cause for this abnormality was identified. The results of this study suggest that eastern box turtles may harbor BTAdV infections at low levels and that infection is rarely associated with clinical disease, potentially identifying BTAdV as a host-adapted pathogen. Future studies should focus on this pathogen's ability to induce clinical disease and its potential impact on recovery efforts for this species.


Assuntos
Infecções por Adenoviridae/veterinária , Adenoviridae/classificação , Adenoviridae/isolamento & purificação , Tartarugas/virologia , Infecções por Adenoviridae/epidemiologia , Infecções por Adenoviridae/virologia , Animais , Animais Selvagens , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Masculino , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Prevalência , Sensibilidade e Especificidade , Virginia/epidemiologia
2.
Oral Dis ; 26(1): 72-80, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31670871

RESUMO

OBJECTIVES: The objective of this study was to report the integrated observations of high-risk HPV-related oral squamous carcinoma (OSCC) at our national referral center for cancer, the Dharmais National Cancer Hospital (DNCH), Jakarta, from 2003 to 2013. MATERIALS AND METHODS: Seventy-eight formalin-fixed paraffin-embedded specimens obtained from OSCC cases were collected from 2003 to 2013 DNCH archives and were included in this high-risk HPV (HR-HPV) study. Seventy-nine DNA samples from the normal oral mucosa of healthy individuals were obtained from the Oral Biology Laboratory DNA archives from 2001 to 2005. Glyceraldehyde 3-phosphate dehydrogenase was used as a control to ensure the DNA integrity for the subsequent HPV DNA PCR detection. High-risk HPV16/18 DNA amplification was conducted by nested PCR using two pairs of primers that were designed specifically to identify the region of gene L1 HPV16 and the HPV16/18 region. RESULTS AND CONCLUSIONS: A high prevalence of HPV16/18 was detected in OSCC cases (17.9%). HPV18 occurred more often than HPV16 (86%) among OSCC patients who were HPV positive. This result supports high HPV18 prevalence among Indonesian cervical cancer patients studied in 1995 and 2006. The prevalence of high-risk HPV remains low in the normal Indonesian population (3.8%), but HPV16 is consistently more frequently detected in non-cancer populations.


Assuntos
Carcinoma de Células Escamosas/virologia , Neoplasias Bucais/virologia , Infecções por Papillomavirus/epidemiologia , Adulto , Carcinoma de Células Escamosas/epidemiologia , DNA Viral/isolamento & purificação , Feminino , Técnicas de Genotipagem , Papillomavirus Humano 16/isolamento & purificação , Papillomavirus Humano 18/isolamento & purificação , Humanos , Indonésia/epidemiologia , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/epidemiologia , Infecções por Papillomavirus/complicações , Prevalência
3.
Int J Cancer ; 146(3): 810-818, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30980692

RESUMO

Primary prevention through the use of human papillomavirus (HPV) vaccination is expected to impact both cervical intraepithelial neoplasia (CIN) and adenocarcinoma in situ (AIS). While CIN is well described, less is known about the epidemiology of AIS, a rare cervical precancer. We identified AIS and CIN grade 3 (CIN3) cases through population-based surveillance, and analyzed data on HPV types and incidence trends overall, and among women screened for cervical cancer. From 2008 to 2015, 470 AIS and 6,587 CIN3 cases were identified. The median age of women with AIS was older than those with CIN3 (35 vs. 31 years; p < 0.01). HPV16 was the most frequently detected type in both AIS and CIN3 (57% in AIS; 58% in CIN3), whereas HPV18 was the second most common type in AIS and less common in CIN3 (38% vs. 5%; p < 0.01). AIS lesions were more likely than CIN3 lesions to be positive for high-risk types targeted by the bivalent and quadrivalent vaccines (HPV16/18, 92% vs. 63%; p < 0.01), and 9-valent vaccine (HPV16/18/31/33/45/52/58, 95% vs. 87%; p < 0.01). AIS incidence rates decreased significantly in the 21-24 year age group (annual percent change [APC] overall: -22.1%, 95% CI: -33.9 to -8.2; APC among screened: -16.1%, 95% CI: -28.8 to -1.2), but did not decrease significantly in any older age group. This report on the largest number of genotyped AIS cases to date suggests an important opportunity for vaccine prevention of AIS, and is the first to document a decline in AIS incidence rates among young women during the vaccine era.


Assuntos
Adenocarcinoma in Situ/epidemiologia , Neoplasia Intraepitelial Cervical/epidemiologia , Papillomavirus Humano 16/isolamento & purificação , Papillomavirus Humano 18/isolamento & purificação , Infecções por Papillomavirus/epidemiologia , Lesões Pré-Cancerosas/epidemiologia , Adenocarcinoma in Situ/prevenção & controle , Adenocarcinoma in Situ/virologia , Adolescente , Adulto , Fatores Etários , Neoplasia Intraepitelial Cervical/prevenção & controle , Neoplasia Intraepitelial Cervical/virologia , DNA Viral/isolamento & purificação , Feminino , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , Humanos , Incidência , Programas de Rastreamento/estatística & dados numéricos , Infecções por Papillomavirus/prevenção & controle , Infecções por Papillomavirus/virologia , Vacinas contra Papillomavirus/uso terapêutico , Lesões Pré-Cancerosas/prevenção & controle , Lesões Pré-Cancerosas/virologia , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/prevenção & controle , Neoplasias do Colo do Útero/virologia , Adulto Jovem
4.
J Water Health ; 17(6): 971-977, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31850903

RESUMO

Torque teno virus (TTV) is a single-stranded DNA virus which is predominantly transmitted by the fecal-oral route and may be excreted in the absence of the clinical symptoms. TTV was previously considered a probable cause of hepatitis, but further studies could not strongly connect TTV to any serious health problem. TTV is highly resistant to water and wastewater treatment processes and can be a useful indicator for determining the fecal contamination of water. The purpose of the present study was to assess the prevalence and molecular characterization of TTV in treated wastewater in Tehran. Thirteen effluent samples were collected monthly from the biggest wastewater treatment plant in Tehran, Iran (from September 2017 to August 2018). The presence of the TTV was monitored in the samples by the nested polymerase chain reaction (PCR) method. The TTV genome was found in 76.9% of the samples, and TTV of groups 1 and 3 were determined using phylogenetic analysis. Therefore, treated wastewater can play a key role in the transmission of TTV and the usage of treated wastewater as a source of potable water needs to be carefully controlled.


Assuntos
DNA Viral/genética , Torque teno virus/isolamento & purificação , Águas Residuárias/virologia , DNA Viral/isolamento & purificação , Humanos , Irã (Geográfico) , Filogenia , Reação em Cadeia da Polimerase/métodos , Prevalência , Torque teno virus/genética
5.
Braz Oral Res ; 33: e091, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31778470

RESUMO

Human Papillomavirus (HPV) has considerable tropism for epithelial and mucosal tissues and can therefore be found in several anatomical sites, including the oral cavity. This study aimed to investigate the presence of HPV-DNA and the most frequent viral types in patients using full dentures, compare to patients not using full dentures and to associate its presence with socio-epidemiological and behavioral factors. The study consisted of 90 patients with or without full dentures at the time of collection, treated at a public dental clinic. The samples were obtained by exfoliating the oral cavity, and analyzed for HPV-DNA using the nested PCR with PGMY09/11 (450-bp), and general primers GP5+/GP6+ (150-bp). Genotyping was performed by specific-type PCR to HPV 6, 11, 16, 18, 31, 33, and 45; and Restriction Fragment Length Polymorphism (RFLP). Pearson's Chi-square test (x 2 ) or Fisher's exact test were applied and significant variables in these tests were analyzed by multinomial logistic regression analysis to estimate odds ratio (OR). HPV-DNA was detected in 27.7% of samples and, among those obtained from patients using full dentures, positivity for HPV-DNA was 41.9% (p = 0.025). The most frequent viral types were low-risk HPV 6 and 11, and high-risk HPV 31 and 45. Patients who used full dentures had an odds ratio of 2.1 to be positive for HPV DNA. Our results indicate the need for periodic dental follow-up of patients with full dentures in order to preserve the basic conditions of oral health, and also to monitor the appearance of lesions with malignant potential.


Assuntos
Prótese Total/virologia , Vírus Oncogênicos/isolamento & purificação , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/virologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil/epidemiologia , Carcinogênese , Estudos Transversais , DNA Viral/isolamento & purificação , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/virologia , Infecções por Papillomavirus/epidemiologia , Reação em Cadeia da Polimerase , Prevalência , Fatores de Risco , Fatores Socioeconômicos
6.
Parasit Vectors ; 12(1): 464, 2019 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-31585545

RESUMO

BACKGROUND: Bluetongue disease of ruminants is a typical insect-borne disease caused by bluetongue virus (BTV) of the genus Orbivirus (family Reoviridae) and transmitted by some species of Culicoides (Diptera: Ceratopogonidae). Recently, the detection of BTV in yaks in high altitude meadows of the Shangri-La district of Yunnan Province, China, prompted an investigation of the Culicoides fauna as potential vectors of BTV. METHODS: A total of 806 Culicoides midges were collected by light trapping at three sites at altitudes ranging from 1800 to 3300 m. The species were identified based on morphology and the DNA sequences of cytochrome c oxidase subunit 1 (cox1). PCR and quantitative PCR following reverse transcription were used to test for the presence of BTV RNA in Culicoides spp. A phylogenetic analysis was used to analyze the cox1 sequences of some specimens. RESULTS: Four species dominated these collections and cox1 barcoding revealed that at least two of these appear to belong to species new to science. Culicoides tainanus and a cryptic species morphologically similar to C. tainanus dominated low altitude valley collections while C. nielamensis was the most abundant species in the high-altitude meadow. A species related to C. obsoletus occurred at all altitudes but did not dominate any of the collections. BTV RT-qPCR analysis detected BTV RNA in two specimens of C. tainanus, in one specimen closely related to C. tainanus and in one specimen closely related to C. obsoletus by barcode sequencing. CONCLUSIONS: This study suggests that BTV in high altitude areas of Yunnan is being transmitted by three species of Culicoides, two of which appear to be new to science. This research may be useful in improving understanding of the effects of global warming on arboviral disease epidemiology and further study is important in research into disease control and prevention.


Assuntos
Bluetongue/transmissão , Doenças dos Bovinos/transmissão , Ceratopogonidae/virologia , Insetos Vetores/virologia , Altitude , Animais , Sequência de Bases , Bluetongue/epidemiologia , Vírus Bluetongue/classificação , Vírus Bluetongue/genética , Vírus Bluetongue/isolamento & purificação , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/virologia , Ceratopogonidae/classificação , China/epidemiologia , Código de Barras de DNA Taxonômico/veterinária , DNA Viral/química , DNA Viral/isolamento & purificação , Complexo IV da Cadeia de Transporte de Elétrons/genética , Cabras , Insetos Vetores/classificação , Filogenia , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Transcrição Reversa , Ruminantes , Estudos Soroepidemiológicos
7.
Vet Microbiol ; 237: 108370, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31585643

RESUMO

Caprine alphaherpesvirus 1 (CpHV-1) is a pathogen associated with systemic infection and respiratory disease in kids and subclinical infection or reproductive failure and abortions in adult goats. The enzyme thymidine kinase (TK) is an important viral product involved in nucleotide synthesis. This property makes the tk gene a common target for herpesvirus attenuation. Here we deleted the tk gene of a CpHV-1 isolate and characterized the recombinant CpHV-1ΔTKin vitro and in vivo. In vitro characterization revealed that the recombinant CpHV-1ΔTK replicated to similar titers and produced plaques of similar size to the parental CpHV-1 strain in BT and CRIB cell lines. Upon intranasal inoculation of young goats, the parental virus replicated more efficiently and for a longer period than the recombinant virus. In addition, infection with the parental virus resulted in mild systemic and respiratory signs whereas the kids inoculated with the recombinant CpHV-1ΔTK virus remained healthy. Goats inoculated with the parental virus also developed higher neutralizing antibody titers when compared to CpHV-1ΔTK inoculated animals. Dexamethasone (Dx) administration on days 35-39 post-inoculation did not result in virus shedding in nasal secretions, indicating lack of reactivation from latency. However, viral DNA was detected in the trigeminal ganglia of animals euthanized at 14 days post-Dx, indicating that both viruses successfully established latent infection. Our results show that the recombinant CpHV-1ΔTK presents an attenuated phenotype when compared to the parental virus, and hence may represent a promising vaccine candidate to prevent CpHV-1 disease in goats.


Assuntos
Alphaherpesvirinae/genética , Deleção de Genes , Doenças das Cabras/virologia , Timidina Quinase/genética , Alphaherpesvirinae/patogenicidade , Animais , Bovinos , Linhagem Celular , DNA Viral/isolamento & purificação , Dexametasona/farmacologia , Regulação Enzimológica da Expressão Gênica , Regulação Viral da Expressão Gênica , Cabras , Muco/virologia , Proteínas Virais , Eliminação de Partículas Virais
8.
BMC Infect Dis ; 19(1): 773, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31484497

RESUMO

BACKGROUND: The etiology of acute liver failure (ALF) is often unknown and reported to be associated with herpesviruses in a number of cases. In this study, we examined for betaherpesviruses infections in patients with ALF of unknown etiology using a multiplex qPCR to Betaherpesviruses subfamily. METHODS: Liver explant and serum samples from 27 patients with ALF of unknown etiology were analyzed with the aid of multiplex qPCR to identify betaherpesviruses. All positive samples were sequenced to confirm herpes infection and liver enzyme levels evaluated. RESULTS: Betaherpesviruses infection was effectively detected using multiplex qPCR. Six (22%) HHV-6, one (3%) HCMV and two (7%) dual infections (one with HHV-7/HHV-6, and the other with HHV-7/ HCMV). Interestingly, HHV-7 was only detected in the presence of other betaherpesviruses. Sequencing information confirmed betaherpesviruses infection. High hepatic enzyme levels and INR values> 1.5 were determined in all betaherpesvirus-positive patients. CONCLUSIONS: Multiplex qPCR facilitated efficient quantification, indicating that differentiation between betaherpesviruses is possible with the sole use of real-time PCR. Liver explant and serum samples were positive for some betaherpesviruses, and coinfection of HHV-7 with HHV-6 and HCMV was additionally detected. Based on these results, we propose that ALF patients should be screened for the presence of betaherpesviruses.


Assuntos
Betaherpesvirinae/genética , Betaherpesvirinae/isolamento & purificação , Infecções por Herpesviridae/diagnóstico , Falência Hepática Aguda/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real , Adolescente , Adulto , Brasil/epidemiologia , Criança , DNA Viral/sangue , DNA Viral/isolamento & purificação , Diagnóstico Diferencial , Feminino , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/virologia , Humanos , Incidência , Falência Hepática Aguda/epidemiologia , Falência Hepática Aguda/virologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real/métodos , Adulto Jovem
9.
Biosens Bioelectron ; 143: 111610, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31445386

RESUMO

A label-free and efficient electrochemical biosensor was developed for the ultrasensitive detection of EBV-related DNA by combing AgDNCs@DNA/AgNCs nanocomposites with noncanonical lambda exonuclease (λ exo)-assisted target recycling (LNTR). The conjugates of AgDNCs, DNA/AgNCs and probe DNA (pDNA-AgDNCs@DNA/AgNCs conjugates) worked as not only ideal nanocarriers but also efficient electrochemical tags. LNTR didn't require phosphorylated substrates and could be triggered specifically by target DNA, leading to the recycling use of target DNA and the liberation of plentiful linker probes (LP). Subsequently, the LP hybridized with the capture probes on the electrode and then bond to pDNA-AgDNCs@DNA/AgNCs conjugates, generating a sensitive electric signal directly. What's more, the signal amplification effects of DNA/AgNCs and LNTR were investigated. Under the optimal conditions, the proposed method exhibited a wide linear range of 1 fM to 1 nM and the detection limit down to 0.38 fM. In addition, the developed biosensing method exhibited excellent specificity and was successfully applied to detect target DNA in complex biological matrix. The proposed biosensor without extra bio-labels may provide a promising platform in bioanalysis and biochemical research.


Assuntos
Técnicas Biossensoriais , DNA Viral/isolamento & purificação , Técnicas Eletroquímicas , Herpesvirus Humano 4/isolamento & purificação , DNA Viral/química , Exonucleases/química , Herpesvirus Humano 4/química , Humanos , Nanocompostos/química
10.
BMC Infect Dis ; 19(1): 624, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31307413

RESUMO

BACKGROUND: Two outbreaks of epidemic keratoconjunctivitis (EKC) occurred successively with an interval of 5 days in two primary boarding schools in Weixi Lisu Autonomous County, Diqing, and Tibetan Autonomous Prefecture, Yunnan. The aims of this study were to determine the intensity and characteristics of the outbreaks, as well as the clinical manifestations in the patients, the risk factors for infection and the pathogen responsible for the two outbreaks. METHODS: An outbreak investigation was conducted in two primary schools, and a case-control study including patients from the Weixi County Ethnic Primary School was performed. Relevant specimens were collected according to the case definition, and next-generation sequencing was employed to identify the pathogen. An epidemiological investigation method was used to analyse the related epidemiological characteristics, such as risk factors. The phylogenetic tree was constructed by MEGA 7.0. RESULTS: A total of 331 acute conjunctivitis cases, including probable cases of EKC, were reported in the two schools, and the attack rates were 30.59% (171/559, 95%CI: 26.76-34.42) and 20.41% (160/784, 95%CI: 17.58-23.24), respectively. Cases occurred in all grades and classes in both schools, and only one staff member in each school presented illness. The epidemics lasted for 54 days and 45 days, respectively. The patients had typical manifestations of EKC, such as acute onset, follicular hyperplasia, pseudomembrane formation, preauricular lymphadenopathy, corneal involvement and blurred vision, and a relatively long disease course (average 9.40 days, longest 23 days and shortest 7 days). The risk factor for infection was close contact with a patient or personal items contaminated by a patient. The pathogen responsible for the outbreaks was HAdV-8. The virus was highly similar to the 2016 HAdV-8 strain from Tibet, China. CONCLUSIONS: This study strongly suggests that HAdV-8 could lead to serious consequences. This is the second report of a HAdV-8-associated EKC outbreak in mainland China. Tibetan HAdV-8 might be circulating in southwest China; therefore, it is necessary to monitor the pathogens causing acute conjunctivitis in this area.


Assuntos
Infecções por Adenovirus Humanos/diagnóstico , Adenovírus Humanos/isolamento & purificação , Ceratoconjuntivite/diagnóstico , Infecções por Adenovirus Humanos/epidemiologia , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/classificação , Adenovírus Humanos/genética , Adolescente , Adulto , Estudos de Casos e Controles , Criança , China/epidemiologia , DNA Viral/isolamento & purificação , DNA Viral/metabolismo , Surtos de Doenças , Feminino , Humanos , Ceratoconjuntivite/epidemiologia , Ceratoconjuntivite/virologia , Masculino , Filogenia , Fatores de Risco , Instituições Acadêmicas , Análise de Sequência de DNA , Adulto Jovem
11.
Taiwan J Obstet Gynecol ; 58(4): 477-481, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31307736

RESUMO

OBJECTIVE: to correlate the detection rate of high risk HPV (HR-HPV) DNA between self-collected and clinician-collected testing. MATERIALS AND METHODS: A cross-sectional analytic study was conducted in 400 women undergoing cervical cancer screening program during February and May 2015. The procedure began with self-collected method and then clinician-collected method. Then, the specimens were processed and interpreted with the same technique. If the results from either methods were positive for HPV genotype 16 or 18, colposcopy was performed. We also conducted cytology testing for the participants. If the results were abnormal (ASC-US+), colposcopy was also performed. RESULTS: The detection rate of HR-HPV DNA was 10.0% and 7.5% by self-collected and clinician-collected specimen, respectively (kappa = 0.73). HR-HPV positive rate in cytology ASC-US+ was no significantly different between groups. HR-HPV DNAs were positive in every HSIL (100% detection rate). HPV DNA test positive for detection CIN+ was not significantly different between self-collected and clinician-collected testing. CONCLUSION: self-collected HPV testing can be used as an alternative option for primary cervical screening program. Detection rate of high grade lesion is similar to clinician-collected test.


Assuntos
Neoplasia Intraepitelial Cervical/patologia , Detecção Precoce de Câncer/métodos , Papillomavirus Humano 16/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Manejo de Espécimes/métodos , Neoplasias do Colo do Útero/patologia , Adulto , Neoplasia Intraepitelial Cervical/cirurgia , Neoplasia Intraepitelial Cervical/virologia , Colposcopia/métodos , Estudos Transversais , Citodiagnóstico/métodos , DNA Viral/isolamento & purificação , Feminino , Humanos , Pessoa de Meia-Idade , Medição de Risco , Autoexame/métodos , Sensibilidade e Especificidade , Tailândia , Neoplasias do Colo do Útero/cirurgia , Neoplasias do Colo do Útero/virologia
12.
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
13.
Artigo em Inglês | MEDLINE | ID: mdl-31203107

RESUMO

Adeno-associated virus (AAV) is a strong candidate for single-gene mutation gene therapy. AAV comes in several serotypes that target different organs in the body. The current purification methods for AAV vectors often rely on serotype dependent affinity chromatography. However, it is desired to create a platform for AAV purification that mirrors the evolution of antibody platform processes. To do this, any serotype dependent steps need to be removed from the process. The harvest and initial capture steps that can satisfy all of the needs of a platform AAV process is the use of low pH and Triton in the harvest, followed by filtration and cation exchange chromatography (CEX) for initial capture. The low pH hydrolyses and removes the host cell DNA, a difficult contaminate to remove. CEX then provides a concentration and capture step. The only step that remains is to determine the polishing and final formulation. This harvest strategy provides a serotype independent purification that removes both host cell DNA and host cell proteins and is friendly to scale-up for future AAV processes.


Assuntos
Cromatografia por Troca Iônica/métodos , DNA Viral/isolamento & purificação , Dependovirus/química , DNA Viral/química , DNA Viral/genética , Dependovirus/genética , Vetores Genéticos/química , Vetores Genéticos/genética , Concentração de Íons de Hidrogênio
14.
Eur J Obstet Gynecol Reprod Biol ; 239: 11-15, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31158788

RESUMO

OBJECTIVE: To determine the relationship between the presence of detectable HBV DNA in the follicular fluid in HBV carriers with IVF/ICSI treatment outcome. STUDY DESIGN: A prospective observational study conducted in the Assisted Reproductive Unit, a tertiary referral centre affiliated with the Department of Obstetrics and Gynecology, The Chinese University of Hong Kong; and the Union Reproductive Medicine Centre at Union Hospital, Hong Kong. The primary outcome measure was pregnancy rate. Secondary outcome measures were the prevalence of detectable HBV DNA in the follicular fluid, implantation rate, clinical pregnancy rate, ongoing pregnancy rate and live birth rate. RESULTS: HBV DNA was detected in the follicular fluid of 28 (43.8%) of the 64 women, and the mean level in this group in log10 copies/mL (±SD) was 4.36 ± 1.85. Women with detectable follicular fluid HBV DNA were younger, lighter, had longer duration of infertility, higher incidence of detectable serum HBV DNA (OR 4.592, 95% C I 2.333-9.038), and significantly wider range in the number of total fertilized, viable embryos, and blastocyst rate, but no difference in cycle characteristics, stimulation and pregnancy outcomes, although the almost doubled ongoing pregnancy/live birth rate per cycle initiated (60.7% versus 38.9%) failed to reach statistical significance due to the small numbers. CONCLUSION: Our results suggested HBV infection did not appear to be detrimental to the outcome of IVF/ICSI treatment.


Assuntos
DNA Viral/isolamento & purificação , Líquido Folicular/virologia , Hepatite B/complicações , Infertilidade Feminina/virologia , Injeções de Esperma Intracitoplásmicas/estatística & dados numéricos , Adulto , Feminino , Hepatite B/virologia , Humanos , Gravidez , Taxa de Gravidez , Estudos Prospectivos
15.
Indian J Med Res ; 149(3): 389-395, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-31249205

RESUMO

Background & objectives: : Nucleic acid amplification test (NAT) in blood donor screening not only detects window period (WP) donors but also those with chronic occult infections which are negative by routine serological screening. This study was conducted to determine the time trend of NAT positivity and seroprevalence of transfusion-transmitted infections (TTIs) through a period of six years and evaluate the strength of NAT as a supplementary test in identifying the cryptic carriers in blood donor population. Methods: : A total of 1,01,411 blood donations were screened between January 2011 and December 2016 by the ELISA and individual donor (ID) NAT Procleix Ultrio Plus Assay. Additional molecular and serological assays were done on the NAT yield samples to differentiate the type of cryptic carriers. Results: : NAT yields comprised 0.05 per cent (50/101411) of the total samples tested with a yield rate of 1/2028. Hepatitis B virus (HBV) contributed to 80 per cent of the total NAT yields and the rest 20 per cent due to hepatitis C virus (HCV). Majority of HBV NAT yields (75%) were from chronic occult donors and 25 per cent were WP donors. Both HBV and HCV NAT yields had a wide range of viral count. There was no HIV NAT yield. A significant decline in the prevalence rate of TTIs through the study period of six years was observed. Interpretation & conclusions: : The cryptic infections found in blood donors increase the risk of TTIs. Blood screening by both serology and NAT can reduce this threat.


Assuntos
Antígenos de Superfície da Hepatite B/sangue , Técnicas de Amplificação de Ácido Nucleico/métodos , Estudos Soroepidemiológicos , Reação Transfusional/sangue , Adulto , Grupo com Ancestrais do Continente Asiático , Doadores de Sangue , Segurança do Sangue , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Hepacivirus/genética , Hepacivirus/isolamento & purificação , Hepacivirus/patogenicidade , Hepatite B/sangue , Hepatite B/epidemiologia , Hepatite B/virologia , Antígenos de Superfície da Hepatite B/isolamento & purificação , Vírus da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , Vírus da Hepatite B/patogenicidade , Hepatite C/sangue , Hepatite C/epidemiologia , Hepatite C/virologia , Humanos , Índia/epidemiologia , Masculino , Reação Transfusional/epidemiologia , Reação Transfusional/genética , Reação Transfusional/virologia
16.
ScientificWorldJournal ; 2019: 2561530, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31061653

RESUMO

Fewer studies have been done over the years to establish the association of human papillomavirus (HPV) with head and neck squamous cell carcinoma (HNSSC) within the subregions of sub-Saharan Africa, and thus this study was designed to investigate the presence of HPV in HNSCC at a tertiary hospital in Ghana, providing additional evidence on the need to explore similar studies in other subregions. A retrospective cross-sectional study was employed to investigate the presence of the DNA of HPV genotypes in HNSCC archived tissue. A total of 100 HNSCC cases were classified as suitable for HPV genotyping. HPV-DNA was detected in 18% of the HNSCC cases, with 17 being HPV-16 and 1 dual infection with HPV-16 and HPV-18. HPV was prevalent in 50% of oropharyngeal cancers, 27% of laryngeal cancers, and 23% of oral cavity cancers. HPV E6/E7 oncogenic DNA was found in 18% of the HNSCC cases, with HPV-16 being the predominant genotype present. The pattern of HPV association was similar to earlier reported studies, recording a higher prevalence in oropharyngeal cancers, followed by laryngeal cancers and oral cavity cancers.


Assuntos
Papillomaviridae/isolamento & purificação , Carcinoma de Células Escamosas de Cabeça e Pescoço/virologia , Adolescente , Adulto , África ao Sul do Saara , Idoso , Idoso de 80 Anos ou mais , Criança , Estudos Transversais , DNA Viral/isolamento & purificação , Feminino , Genótipo , Gana , Humanos , Masculino , Pessoa de Meia-Idade , Papillomaviridae/genética , Estudos Retrospectivos , Centros de Atenção Terciária
17.
PLoS One ; 14(5): e0215945, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31042762

RESUMO

The composition of the vaginal microbiome, including both the presence of pathogens involved in sexually transmitted infections (STI) as well as commensal microbiota, has been shown to have important associations for a woman's reproductive and general health. Currently, healthcare providers cannot offer comprehensive vaginal microbiome screening, but are limited to the detection of individual pathogens, such as high-risk human papillomavirus (hrHPV), the predominant cause of cervical cancer. There is no single test on the market that combines HPV, STI, and microbiome screening. Here, we describe a novel inclusive vaginal health assay that combines self-sampling with sequencing-based HPV detection and genotyping, vaginal microbiome analysis, and STI-associated pathogen detection. The assay includes genotyping and detection of 14 hrHPV types, 5 low-risk HPV types (lrHPV), as well as the relative abundance of 31 bacterial taxa of clinical importance, including Lactobacillus, Sneathia, Gardnerella, and 3 pathogens involved in STI, with high sensitivity, specificity, and reproducibility. For each of these taxa, reference ranges were determined in a group of 50 self-reported healthy women. The HPV sequencing portion of the test was evaluated against the digene High-Risk HPV HC2 DNA test. For hrHPV genotyping, agreement was 95.3% with a kappa of 0.804 (601 samples); after removal of samples in which the digene hrHPV probe showed cross-reactivity with lrHPV types, the sensitivity and specificity of the hrHPV genotyping assay were 94.5% and 96.6%, respectively, with a kappa of 0.841. For lrHPV genotyping, agreement was 93.9% with a kappa of 0.788 (148 samples), while sensitivity and specificity were 100% and 92.9%, respectively. This novel assay could be used to complement conventional cervical cancer screening, because its self-sampling format can expand access among women who would otherwise not participate, and because of its additional information about the composition of the vaginal microbiome and the presence of pathogens.


Assuntos
Microbiota , Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Doenças Sexualmente Transmissíveis/diagnóstico , Vagina/virologia , Adolescente , Adulto , Proteínas do Capsídeo/genética , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Gardnerella/genética , Gardnerella/isolamento & purificação , Genótipo , Humanos , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Limite de Detecção , Pessoa de Meia-Idade , Proteínas Oncogênicas Virais/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/virologia , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Doenças Sexualmente Transmissíveis/virologia , Vagina/microbiologia , Adulto Jovem
18.
Microb Pathog ; 132: 162-165, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31054871

RESUMO

Head and neck cancers (HNCs) are a major health problem and a leading cause of morbidity and mortality worldwide. More than 90% of these tumours are head and neck squamous cell carcinomas (HNSCCs). Amongst the common risk factors for HNCs (tobacco and alcohol use), there is a strong association of human papillomavirus (HPV) with HNSCCs. HPV type 16 (HPV 16), the major high-risk HPV type, is most commonly associated with HPV-driven HNSCCs. The promiscuous nature of the major HPV oncogene, E7, allows its interaction with a myriad of host proteins including STING, a component of the viral DNA-sensing cyclic GMP-AMP synthase (cGAS) - stimulator of interferon genes (STING) machinery. Sensing of viral DNA by the cGAS-STING machinery results in a type I interferon (IFN)-mediated anti-viral response. Amelioration of IFN responses resulting from the direct blockade of STING by E7 was first demonstrated in high-risk HPV type 18 (HPV 18) positive (+) cervical squamous cell carcinoma (CESC) cells. However, the role of E7 from HPV 16 (HPV 16E7) in antagonising cGAS-STING responses have not been investigated, let alone in the context of HNSCCs. Here, we show that HPV 16E7+, but not HPV 16E7 negative (-), HNSCC cells respond poorly to cGAS-STING activation stimulus. We further confirm that this inhibition occurred via the highly conserved LXCXE motif in 16E7. This finding contributes to the better understanding of role of high-risk HPV E7 in blocking cGAS-STING pathway, especially in the context of HNSCCs.


Assuntos
DNA Viral/isolamento & purificação , Neoplasias de Cabeça e Pescoço/virologia , Papillomavirus Humano 16/genética , Nucleotidiltransferases/genética , Infecções por Papillomavirus/virologia , Carcinoma de Células Escamosas de Cabeça e Pescoço/virologia , Linhagem Celular Tumoral , DNA Viral/genética , Regulação Viral da Expressão Gênica , Neoplasias de Cabeça e Pescoço/complicações , Papillomavirus Humano 16/metabolismo , Humanos , Interferon Tipo I/genética , Interferon Tipo I/metabolismo , Nucleotidiltransferases/antagonistas & inibidores , Nucleotidiltransferases/metabolismo , Proteínas E7 de Papillomavirus/genética , Proteínas E7 de Papillomavirus/metabolismo , Infecções por Papillomavirus/complicações , Carcinoma de Células Escamosas de Cabeça e Pescoço/complicações
19.
Gynecol Oncol ; 154(1): 110-117, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30982556

RESUMO

OBJECTIVE: Nine-valent human papillomavirus (9vHPV) vaccine efficacy against disease and cervical surgeries related to all nine vaccine components was assessed compared with a historic placebo population. This was not assessed in the 9vHPV vaccine efficacy trial since the trial was quadrivalent HPV (qHPV) vaccine-controlled, efficacy was measured for the five HPV types covered only by 9vHPV vaccine (HPV31/33/45/52/58), but not the four types covered by both vaccines (HPV6/11/16/18). METHODS: Three international, randomized, double-blind studies were conducted using the same methodology. In the 9vHPV vaccine study (NCT00543543), 7106 and 7109 women received 9vHPV or qHPV vaccine, respectively. In the historic qHPV vaccine studies (FUTURE I [NCT00092521] and II [NCT00092534]), 8810 and 8812 women received qHPV vaccine or placebo, respectively, based on the same eligibility criteria. Cervical cytological testing was performed regularly. Biopsy or definitive therapy specimens were assessed for HPV DNA. RESULTS: Among women negative for 14 HPV types prior to vaccination, incidence of high-grade cervical disease (9vHPV, n = 2 cases; placebo, n = 141 cases) and cervical surgery (9vHPV, n = 3 cases; placebo, n = 170 cases) related to the nine HPV types was reduced by 98.2% (95% confidence interval [CI], 93.6-99.7) and 97.8% (95% CI, 93.4-99.4), respectively. The 9vHPV vaccine did not prevent disease related to vaccine HPV types detected at baseline, but significantly reduced cervical, vulvar, and vaginal diseases related to other vaccine HPV types. CONCLUSIONS: Effective implementation of the 9vHPV vaccine may substantially reduce the burden of HPV-related diseases and related medical procedures. TRIAL REGISTRATIONS: clinicaltrials.gov: NCT00543543, NCT00092521, NCT00092534.


Assuntos
Infecções por Papillomavirus/prevenção & controle , Vacinas contra Papillomavirus/administração & dosagem , Displasia do Colo do Útero/prevenção & controle , Doenças Vaginais/prevenção & controle , Doenças da Vulva/prevenção & controle , Adulto , DNA Viral/isolamento & purificação , Método Duplo-Cego , Feminino , Humanos , Papillomaviridae/genética , Papillomaviridae/imunologia , Infecções por Papillomavirus/patologia , Vacinas contra Papillomavirus/efeitos adversos , Vacinas contra Papillomavirus/imunologia , Displasia do Colo do Útero/patologia , Displasia do Colo do Útero/cirurgia , Displasia do Colo do Útero/virologia , Doenças Vaginais/patologia , Doenças Vaginais/virologia , Doenças da Vulva/patologia , Doenças da Vulva/virologia , Adulto Jovem
20.
J Low Genit Tract Dis ; 23(3): 200-204, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30946259

RESUMO

OBJECTIVE: The aim of this study was to determine the concordance of self- and clinician-collected anorectal swabs for the detection of human papillomavirus (HPV) DNA in a population of HIV-negative men who have sex with men (MSM). METHODS: This cross-sectional study involved recruitment of HIV-negative MSM in a Midwestern US metropolitan area to collect paired sequential self- and clinician-collected anorectal swabs using illustrated instructions. Swabs were tested for type-specific HPV DNA with a comparison of type-specific HPV categories detected by each method. The sensitivity and specificity of self-collection were calculated assuming clinician collection as the criterion standard. McNemar's test and κ statistics were used to determine percent agreement and concordance of self- and clinician-collected swab results. RESULTS: Seventy-eight participants had paired anorectal swab samples of adequate quality for analyses. The sensitivity and specificity of self-collected swabs for detection of all high-risk HPV DNA types were 69.8% and 91.4%, respectively. Similar degrees of sensitivity and specificity of self-collection were seen for other groups of high-risk HPV types. Percent agreement and κ statistic for self- and clinician-collected swabs for all high-risk HPV types were 80.8% and 0.53, respectively. CONCLUSIONS: Self-collected anorectal swab samples showed lower sensitivity but moderate to high specificity for detection of high-risk and vaccine-preventable HPV types compared with clinician-collected swab samples. Self-collection instructional details and the thoroughness of clinician collection of samples may have impacted sensitivity and specificity, suggesting a need to optimize and standardize instructions.


Assuntos
Doenças do Ânus/diagnóstico , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Autoexame/métodos , Manejo de Espécimes/métodos , Adulto , Estudos Transversais , DNA Viral/isolamento & purificação , Humanos , Masculino , Sensibilidade e Especificidade
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