RESUMO
BACKGROUND: Oropharyngeal squamous cell carcinoma (OPSCC) is dominated by tonsillar and tongue base carcinomas (TSCC/BOTSCC), but there are carcinomas at other sites, such as uvula/soft palate/pharyngeal wall here defined as other OPSCC. Human papillomavirus (HPV) positive TSCC/BOTSCC have favorable outcome, and the TNM-classification separates OPSCC into HPV mediated (p16INK4a overexpressing, p16+) and HPV unrelated OPSCC (p16INK4a non-overexpressing, p16-) cancer, but the prognostic role of p16+ in other OPSCC is unclear. AIMS/OBJECTIVES: This study therefore aimed to further investigate the prognostic role of p16+, presence of HPV DNA, or both combined in other OPSCC. MATERIAL AND METHODS: 195 other OPSCC, from patients diagnosed 2000-2018 were tested for p16, and/or presence of HPV DNA and the data correlated to outcome. RESULTS: Neither overall survival, nor disease free survival correlated to presence of p16+ or HPV DNA in other OPSCC. p16+ and HPV DNA presence were correlated (p < .0001), but the sensitivity of p16 as a surrogate marker for presence of HPV DNA was low (49%). CONCLUSIONS AND SIGNIFICANCE: The data suggest that p16+ (and p16+/HPV DNA) positive other OPSCC should be analyzed cautiously and possibly separately from the HPV mediated OPSCC staging group.
Assuntos
Regulação Viral da Expressão Gênica , Papillomavirus Humano 16/genética , Neoplasias Orofaríngeas/genética , Infecções por Papillomavirus/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Neoplasias da Língua/genética , Proteínas do Core Viral/genética , Idoso , Sondas de DNA de HPV/biossíntese , Sondas de DNA de HPV/genética , DNA Viral/genética , Feminino , Seguimentos , Humanos , Masculino , Neoplasias Orofaríngeas/diagnóstico , Neoplasias Orofaríngeas/virologia , Infecções por Papillomavirus/metabolismo , Infecções por Papillomavirus/virologia , Estudos Retrospectivos , Carcinoma de Células Escamosas de Cabeça e Pescoço/diagnóstico , Carcinoma de Células Escamosas de Cabeça e Pescoço/virologia , Neoplasias da Língua/diagnóstico , Neoplasias da Língua/virologia , Proteínas do Core Viral/biossínteseRESUMO
We describe a new assaying system for the detection and genotyping of human papillomavirus (HPV) based on linear-after-the-exponential-PCR(LATE-PCR) and melting curve analysis. The 23 most prevalent HPV strains (types 6, 11, 16, 18, 31, 33, 35, 39, 42, 45, 51, 52, 53, 56, 58, 59, 66, 68, 70, 73, 81, 82, and 83) are assayed in two sealed reaction tubes within 2 h. Good sensitivity and specificity was evaluated by testing cloned HPV DNA and clinical samples. The detection limit was 5-500 copies/reaction depending on the genotype. No cross-reactivity was observed with the other HPV types that are not covered by our method or pathogens tested which were commonly found in female genital tract. When compared with the HPV GenoArray Diagnostic kit, the results from 1104 clinical samples suggest good overall agreement between the two methods,(98.37%, 95% CI: 97.44%-98.97%) and the kappa value was 0.954. Overall, this new HPV genotyping assay system presents a simple, rapid, universally applicable, sensitive, and highly specific detection methodology that should be useful for HPV detection and genotyping, therefore, is potentially of great value in clinical application.
Assuntos
Testes de DNA para Papilomavírus Humano/métodos , Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Sondas de DNA de HPV/genética , Genótipo , Técnicas de Genotipagem/métodos , Humanos , Técnicas de Diagnóstico Molecular/métodos , Papillomaviridae/patogenicidade , Infecções por Papillomavirus/genética , Reação em Cadeia da Polimerase , Sensibilidade e EspecificidadeRESUMO
A cohort of 388 young men enrolled for military service in the Danish army was established and the participants underwent a clinical examination with human papillomavirus (HPV) testing. In addition, a questionnaire containing questions regarding sociodemographic variables, sexual habits and lifestyle factors was completed. The prevalence of HPV was 33.4% in this cohort of uncircumcised men aged 18-29 years. Multiple HPV types were prevalent with one-third of the HPV-positive men being positive for more than one HPV type. Number of recent sexual partners and infrequent condom use were strong risk factors, particularly in men having multiple HPV types. Our findings re-emphasize the importance of sexual transmission and also point to a role of factors that may be related to individual susceptibility as genital warts, alcohol intake and, to a lesser extent, smoking were strongly associated with having multiple HPV types.
Assuntos
Condiloma Acuminado/epidemiologia , Papillomaviridae/classificação , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/epidemiologia , Pênis/virologia , Adolescente , Adulto , Estudos de Coortes , Condiloma Acuminado/virologia , Sondas de DNA de HPV/genética , Dinamarca/epidemiologia , Humanos , Técnicas Imunoenzimáticas , Masculino , Militares , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase , Prevalência , Análise de Regressão , Fatores de Risco , Fatores Socioeconômicos , Inquéritos e Questionários , Adulto JovemAssuntos
Infecções por Papillomavirus/diagnóstico , Neoplasias do Colo do Útero/diagnóstico , Esfregaço Vaginal/estatística & dados numéricos , Adolescente , Adulto , Fatores Etários , Criança , Análise Custo-Benefício , Sondas de DNA de HPV/análise , Sondas de DNA de HPV/genética , DNA Viral/análise , DNA Viral/genética , Feminino , Educação em Saúde , Papillomavirus Humano 16/genética , Papillomavirus Humano 16/imunologia , Papillomavirus Humano 16/isolamento & purificação , Papillomavirus Humano 18/genética , Papillomavirus Humano 18/imunologia , Papillomavirus Humano 18/isolamento & purificação , Humanos , Programas de Rastreamento/economia , Programas de Rastreamento/tendências , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/prevenção & controle , Infecções por Papillomavirus/virologia , Vacinas contra Papillomavirus/economia , Vacinas contra Papillomavirus/imunologia , Sensibilidade e Especificidade , Fatores de Tempo , Neoplasias do Colo do Útero/epidemiologia , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia , Vacinação , Esfregaço Vaginal/economia , Esfregaço Vaginal/métodos , Adulto JovemRESUMO
The novel HPV 9G DNAChips were developed for the detection and discrimination of the HPV genotypes in the clinical samples. The HPV 9G DNAChip established high SBR of 50-70 and 100% target-specific hybridization after 30min hybridization and 2×2min washing at 25°C. We compared the genotyping results of the 959 HPV positive and 82 HPV negative clinical samples by the HPV 9G DNAChip and the sequencing; the results are in 100% agreement. The HPV 9G DNAChip efficiently discriminate 19 HPV genotypes in the 959 HPV positive clinical samples. The results of HPV 9G DNAChip were 100% identical with the sequencing analysis in the detection and discrimination of HPV genotypes in the HPV negative clinical samples. The high SBR, 100% target-specific hybridization, and 25°C hybridization and washing makes the HPV 9G DNAChip a promising diagnostic tool for the accurate HPV genotyping.
Assuntos
Alphapapillomavirus/genética , Sondas de DNA de HPV/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Infecções por Papillomavirus/diagnóstico , DNA Viral/genética , Feminino , Humanos , Limite de Detecção , Técnicas de Diagnóstico Molecular , Tipagem Molecular , Infecções por Papillomavirus/virologia , Análise de Sequência de DNA , Razão Sinal-RuídoRESUMO
AIM: For full-scale analysis of Human Papillomavirus (HPV) status in humans, two minor groove binder (MGB)-based one-step multiplex real-time PCR systems were developed: one to screen 16 high-risk HPV (HR-HPV) types, and one to screen a broader spectrum of HPV types (common HPV or C-HPV). METHODS: Sensitivity and specificity were evaluated using diluted reference plasmids and 20 control human DNA samples. For clinical evaluation, 510 cervical scrape samples were evaluated. RESULTS: The sensitivity assays revealed that the C-HPV detection system could detect 10 ~ 100 plasmid copies/reaction, while the HR-HPV detection system detected 10 ~ 500 copies. The specificity test revealed that the systems did not yield positive signals from the 20 human genomic DNA samples. Performance was tested on 510 usable clinical samples. The HR-HPV results were compared to those from the Hybrid Capture 2 (HC2) test, which assesses 13 HR-HPV types; the concordance level between the two methods was 90.8% with a kappa value of 0.813. CONCLUSIONS: These results showed that our novel MGB-based one-step multiplex real-time PCR method may be used for the diagnosis and mass screening of HPV in clinical and large-scale epidemiological studies.
Assuntos
Testes de DNA para Papilomavírus Humano/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Colo do Útero/virologia , Sondas de DNA de HPV/genética , DNA Viral/química , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Humanos , Papillomaviridae/classificação , Infecções por Papillomavirus/virologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNARESUMO
Most guidelines currently recommend the testing of human papillomavirus (HPV) in ASCUS cases. The most used method for this purpose is Hybrid Capture II (HCII), but PCR techniques with GP5+/6+ primers can be also applied. Furthermore, the HCII high-risk probe test for detection of HPV shows cross-reactivity with low-risk HPV. Although this cross-reactivity has been studied in screening populations, it has received little attention in ASCUS cases. To compare the performance of the HCII high-risk probe test and PCR for the detection of HPV in ASCUS cases. We randomly selected 83 ASCUS cases that were positive for high-risk HPV by HCII and applied the PCR test using MYO9-11 and GP5+/6+ primers to samples from these cases. Our results show cross-reactivity with low-risk HPV in 25.3% (21/83) of the HCII+ PCR+ cases. Regarding the follow-up our results emphasize the importance of HPV typing, especially for HPV 16 infection. We propose the use of PCR techniques using GP5+/6+ consensus primers for the screening of HPV in ASCUS.
Assuntos
Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/genética , Displasia do Colo do Útero/diagnóstico , Displasia do Colo do Útero/genética , Impressões Digitais de DNA/métodos , Primers do DNA/genética , Sondas de DNA de HPV/genética , Feminino , Humanos , Papillomaviridae/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Esfregaço VaginalRESUMO
A novel microfluidic device with microbeads array was developed and sensitive genotyping of human papillomavirus was demonstrated using a multiple-enzyme labeled oligonucleotide-Au nanoparticle bioconjugate as the detection tool. This method utilizes microbeads as sensing platform that was functionalized with the capture probes and modified electron rich proteins, and uses the horseradish peroxidase (HRP)-functionalized gold nanoparticles as label with a secondary DNA probe. The functionalized microbeads were independently introduced into the arrayed chambers using the loading chip slab. A single channel was used to generate weir structures to confine the microbeads and make the beads array accessible by microfluidics. Through "sandwich" hybridization, the enzyme-functionalized Au nanoparticles labels were brought close to the surface of microbeads. The oxidation of biotin-tyramine by hydrogen peroxide resulted in the deposition of multiple biotin moieties onto the surface of beads. This deposition is markedly increased in the presence of immobilized electron rich proteins. Streptavidin-labeled quantum dots were then allowed to bind to the deposited biotin moieties and displayed the signal. Enhanced detection sensitivity was achieved where the large surface area of Au nanoparticle carriers increased the amount HRP bound per sandwiched hybridization. The on-chip genotyping method could discriminate as low as 1fmol/L (10zmol/chip, SNR>3) synthesized HPV oligonucleotides DNA. The chip-based signal enhancement of the amplified assay resulted in 1000 times higher sensitivity than that of off-chip test. In addition, this on-chip format could discriminate and genotype 10copies/µL HPV genomic DNA using the PCR products. These results demonstrated that this on-chip approach can achieve highly sensitive detection and genotyping of target DNA and can be further developed for detection of disease-related biomolecules at the lowest level at their earliest incidence.
Assuntos
Sondas de DNA de HPV , Técnicas Analíticas Microfluídicas/métodos , Papillomaviridae/genética , Pontos Quânticos , Sequência de Bases , Colo do Útero/virologia , Sondas de DNA de HPV/genética , Desenho de Equipamento , Feminino , Genótipo , Ouro , Humanos , Nanopartículas Metálicas , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/estatística & dados numéricos , Papillomaviridae/classificação , Papillomaviridae/isolamento & purificaçãoRESUMO
BACKGROUND: Human papilloma virus (HPV)-related warts persist, evading host immune surveillance, but sometimes disappear with inflammation. OBJECTIVES: To elucidate the immune evasion mechanisms of HPV, we have examined the density, dynamics, and subsets of dendritic cell (DC) types in non-inflammatory or inflammatory HPV-related skin lesions such as warts and Bowen's disease (HPV-Bowen), and compared the epidermal expression levels of macrophage inflammatory protein (MIP)-3α and E-cadherin. METHODS: The expression of various DC markers, MIP-3α, and E-cadherin in the tissue samples obtained from patients with warts, HPV-Bowen and HPV-unrelated skin diseases was evaluated by immunohistochemistry. MIP-3α gene expression levels were examined in warts and HPV-Bowen by in situ hybridization (ISH) and real-time quantitative polymerase chain reaction (RT-qPCR). RESULTS: The numbers of Langerhans cells (LCs) and the expression levels of MIP-3α and E-cadherin were decreased in non-inflammatory warts and HPV-Bowen, as compared with normal skin. Both epidermal LCs and MIP-3α expression reappeared in inflammatory warts, associated with dermal infiltrates composed of many cytotoxic T cells and various subsets of DCs, while cellular infiltrates in HPV-Bowen contained many B cells and plasma cells with sparse infiltration of DCs. The upregulation of MIP-3α gene expression was confirmed in the inflammatory warts and HPV-Bowen by ISH and RT-qPCR. CONCLUSIONS: The depletion of LCs in the non-inflammatory warts and HPV-Bowen is associated with a down-regulation of expression levels of MIP-3α and E-cadherin in the lesional keratinocytes. MIP-3α expression is upregulated in lesional keratinocytes of inflammatory warts, with the subsequent recruitment of various DC subsets and cytotoxic T cells, whereas plasma cell-rich infiltration was induced in HPV-Bowen.
Assuntos
Células Dendríticas/classificação , Células Dendríticas/imunologia , Infecções por Papillomavirus/imunologia , Dermatopatias Virais/imunologia , Apoptose , Sequência de Bases , Doença de Bowen/genética , Doença de Bowen/imunologia , Doença de Bowen/metabolismo , Doença de Bowen/virologia , Caderinas/genética , Caderinas/metabolismo , Quimiocina CCL20/genética , Quimiocina CCL20/metabolismo , Sondas de DNA de HPV/genética , Humanos , Imuno-Histoquímica , Hibridização In Situ , Queratinócitos/patologia , Ceratose Seborreica/genética , Ceratose Seborreica/imunologia , Ceratose Seborreica/metabolismo , Infecções por Papillomavirus/genética , Infecções por Papillomavirus/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Dermatopatias Virais/genética , Dermatopatias Virais/metabolismo , Verrugas/genética , Verrugas/imunologia , Verrugas/metabolismoRESUMO
INTRODUCTION: Academic research has made a significant advancement in understanding the viral causes of cervical cancer and generating the technology for prevention, both at the primary and secondary levels. Human papillomaviruses (HPVs) have been recognized as the first necessary cause of cervical cancer, the second most common cancer in women worldwide. AREAS COVERED: This paper reviews the epidemiological evidence of the causality of HPV in relation to cervical cancer, other genital tract cancers and some cancers of the oral cavity and oropharynx. The review also covers HPV DNA testing as a screening tool. DNA probes of high-risk HPV types in different formats have been fully validated as primary screening tests, as secondary triage tests and as a prognostic marker following treatment of high grade squamous intraepithelial lesions (HSIL). They consistently showed significant superiority over the conventional Pap smears. Biomarkers of the activation of oncogenes (HPV mRNA, p16 and other) are being tested as screening options to improve in sensitivity and specificity, with promising results. HPV vaccines against the two most common HPV types in cancer have completed their Phase III trials with excellent results in efficacy and safety. Combined strategies of HPV vaccination and HPV-based screening tests could theoretically control cervical cancer in any population in which a large coverage with both preventive options is ensured. Accessibility of developing countries to vaccination and low-cost HPV screening options are the barriers to overcome at present. EXPERT OPINION: This paper provides a synthesis of the evidence available supporting the novel paradigm for cervical cancer prevention that has reached a large consensus within the mainstream HPV and cervical cancer prevention research communities. The available technology for prevention and its developments allows real opportunities for cervical cancer elimination in defined populations to be foreseen.
Assuntos
Programas de Rastreamento , Papillomaviridae , Infecções por Papillomavirus/prevenção & controle , Tecnologia Farmacêutica , Neoplasias do Colo do Útero/prevenção & controle , Ensaios Clínicos como Assunto , Análise Custo-Benefício , Sondas de DNA de HPV/genética , Países Desenvolvidos/estatística & dados numéricos , Países em Desenvolvimento/estatística & dados numéricos , Feminino , Humanos , Masculino , Programas de Rastreamento/economia , Programas de Rastreamento/métodos , Teste de Papanicolaou , Papillomaviridae/genética , Papillomaviridae/imunologia , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/virologia , Vacinas contra Papillomavirus/administração & dosagem , Vacinas contra Papillomavirus/economia , Prevenção Primária/economia , Prevenção Primária/métodos , Prevenção Secundária/economia , Prevenção Secundária/métodos , Neoplasias do Colo do Útero/epidemiologia , Neoplasias do Colo do Útero/virologia , Esfregaço VaginalRESUMO
We have developed a new analytical method to detect multiple DNA simultaneously based on the biobarcoded CdSe/ZnS quantum dot (QD) and magnetic microparticle (MMP). It was demonstrated by using oligonucleotide sequences of 64 bases associated with human papillomavirus 16 and 18 L1 genes (HPV-16 and HPV-18) as model systems. This analytical system involves three types of probes, a MMP probe and two streptavidin-modified QD probes. The MMPs are functionalized with HPV-16 and HPV-18 captures DNA to form MMP probes. The QDs are conjugated with HPV-16 or HPV-18 probe DNA along with FAM- or Rox-labeled random DNA to form HPV-16 and HPV-18 QD probes, respectively. A one-step hybridization reaction was performed by mixing the MMP probes, HPV-16 and HPV-18 target DNA (T-16 and T-18), HPV-16 and HPV-18 QD probes. Afterwards, the hybrid-conjugated microparticles were separated by a magnet and heated to remove the MMPs. Finally, the detections of T-16 and T-18 were done by measuring fluorescence signals of FAM and Rox, respectively. Under the optimum conditions, the fluorescence intensity exhibited a good linear dependence on target DNA concentration in the range from 8 × 10⻹¹ to 8 × 10â»9 M. The detection limit of T-16 is up to 7 × 10⻹¹ M (3σ), and that of T-18 is 6 × 10⻹¹ M. Compared with other biobarcode assay methods, the proposed method that QDs were used as the solid support has some advantages including shorter preparation time of QD probes, faster binding kinetics and shorter analytical time. Besides, it is simple and accurate.
Assuntos
DNA/análise , DNA/genética , Pontos Quânticos , Sequência de Bases , Código de Barras de DNA Taxonômico , Sondas de DNA de HPV/genética , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , Humanos , Concentração de Íons de Hidrogênio , Limite de Detecção , Magnetismo , Hibridização de Ácido Nucleico , Concentração Osmolar , Polimorfismo de Nucleotídeo Único , EstreptavidinaRESUMO
For the detection of DNA hybridization, a new electrochemical biosensor was developed on the basis of the interaction of hematoxylin with 20-mer deoxyoligonucleotides (from human papilloma virus, HPV). The study was performed based on the interaction of hematoxylin with an alkanethiol DNA probe self-assembled gold electrode (ss-DNA/AuE) and its hybridization form (ds-DNA/AuE). The optimum conditions were found for the immobilization of HPV probe on the gold electrode (AuE) surface and its hybridization with the target DNA. Electrochemical detection of the self-assembled DNA and the hybridization process were performed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) over the potential range where the accumulated hematoxylin at the modified electrode was electroactive. Observing a remarkable difference between the voltammetric signals of the hematoxylin obtained from different hybridization samples (non-complementary, mismatch and complementary DNAs), we confirmed the potential of the developed biosensor in detecting and discriminating the target complementary DNA from non-complementary and mismatch oligonucleotides. Under optimum conditions, the electrochemical signal had a linear relationship with the concentration of the target DNA ranging from 12.5 nM to 350.0 nM, and the detection limit was 3.8 nM.
Assuntos
Pareamento Incorreto de Bases/genética , Técnicas Biossensoriais/instrumentação , Condutometria/instrumentação , Sondas de DNA de HPV/genética , DNA Viral/genética , Hematoxilina/química , Análise de Sequência de DNA/instrumentação , DNA Viral/análise , DNA Viral/química , Desenho de Equipamento , Análise de Falha de Equipamento , Marcação de Genes/instrumentação , Hematoxilina/análise , Hibridização In Situ/instrumentação , Coloração e Rotulagem/métodosRESUMO
OBJECTIVE: To use flow cytometry to screen cervical samples for the overexpression of human papillomavirus (HPV) E6 and E7 mRNA and compare the performance of this assay with an HPV DNA array for the detection of high-grade cervical lesions. STUDY DESIGN: Cervical samples were analyzed for HPV DNA by clinical arrays, and the overexpression of E6 and E7 viral oncogenes was monitored using an HPV mRNA detection kit that quantifies the intracellular HPV E6 and E7 mRNA on a cell-by-cell basis. RESULTS: HPV positivity increased with severity of histologic lesions. On the basis of histology-confirmed CIN 2+ cases the specificity of HPV assay was 73.9% (95% CI 66.07, 80.88), whereas it was 39.3% (95% CI 31.85, 47.1) for the DNA assay. CONCLUSION: The HPV assay provides an early predictor of persistent HPV infection and may improve cervical cancer screening by increasing the specificity of detecting high-grade lesions.
Assuntos
Sondas de DNA de HPV/genética , DNA Viral/análise , Citometria de Fluxo/métodos , Proteínas Oncogênicas Virais/genética , Proteínas E7 de Papillomavirus/genética , RNA Mensageiro/análise , Neoplasias do Colo do Útero/virologia , DNA Viral/genética , Feminino , Humanos , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/virologia , RNA Mensageiro/genética , Sensibilidade e Especificidade , Análise de Sequência de DNA , Neoplasias do Colo do Útero/diagnósticoRESUMO
OBJECTIVE: To examine the specificity of human papillomavirus (HPV) E6/E7 mRNA testing for intraepithelial precursor lesions and invasive carcinoma of the uterine cervix in 358 women and compare the results with those of the most widely used DNA technique. STUDY DESIGN: For HPV E6/E7 mRNA testing an amplification assay was used. For DNA determination a hybridization assay was applied. Both techniques were used simultaneously in patients with normal morphology (150), cervical intraepithelial neoplasia (173) and invasive carcinoma of the cervix (35). RESULTS: HPV DNA positivity rates were significantly higher than E6/E7 mRNA in women with normal morphology (21-7%), cervical intraepithelial neoplasia (CIN) 1 and 2 (75-43%), and CIN 3 (93-63%). In invasive cervical carcinoma, both methods tested equally high (94% vs. 97%). Considering that E6/E7 up-regulation represents the initial step in cervical carcinogenesis, it can be assumed that this test allows a more specific detection of lesions with a potential for progression. CONCLUSION: HPV E6/E7 mRNA may serve as a more specific discriminator between transient cervical dysplasias and potentially progressive lesions. Accordingly, testing for high-risk HPV E6/E7 mRNA might reduce the psychologic burden associated with HPV-DNA testing.
Assuntos
DNA Viral/análise , Citometria de Fluxo/métodos , Proteínas Oncogênicas Virais/genética , Proteínas E7 de Papillomavirus/genética , RNA Mensageiro/análise , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/virologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Sondas de DNA de HPV/genética , DNA Viral/genética , Feminino , Humanos , Pessoa de Meia-Idade , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/virologia , RNA Mensageiro/genética , Sensibilidade e Especificidade , Análise de Sequência de DNA , Adulto JovemRESUMO
OBJECTIVE: To evaluate if the cellularity of Hybrid Capture samples (Digene, São Paulo, Brazil) influences the results of HPV-DNA Hybrid Capture tests in men. STUDY DESIGN: We harvested material from penile scrapings for the Hybrid Capture HPV test. This material was then used to make cytologic smears, which we used to evaluate for the presence of nonnucleated squamous cells, nucleated squamous cells and glandular cells. The cellularity of nucleated squamous cells was classified as absent, low, moderate or high. Subsequently, we performed the Hybrid Capture test to identify the low and high risk of HPV and compared these results with the cytologic findings. We used the Fisher and odds ratio tests at CI of 95% to determine statistical significance. RESULTS: Of the 88 tests performed, 65 (74.0%) were negative for HPV-DNA and 23 (26.0%) were positive. Nucleated and nonnucleated squamous cells were absent on nine slides, all of which tested negative for HPV. When only nonnudcleated squamous cells were found, 20% of the cases were positive for HPV-DNA (p < 0.0001; OR = 26.185). The presence of nucleated squamous cells correlated with 33% HPV-DNA positivity (p < 0.0001, OR = 49.05). CONCLUSION: Assessing the presence of non-nucleated and nucleated squamous cells on cytologic smears prior to performing an HPV-DNA test is a useful tool for quality control in penile samples.
Assuntos
DNA Viral/análise , Hibridização de Ácido Nucleico/métodos , Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Doenças do Pênis/diagnóstico , Núcleo Celular/patologia , Núcleo Celular/virologia , Citodiagnóstico/métodos , Sondas de DNA de HPV/genética , Células Epiteliais/patologia , Células Epiteliais/virologia , Humanos , Masculino , Infecções por Papillomavirus/virologia , Doenças do Pênis/virologiaRESUMO
A proof-of-concept of an electrochemical genosensor array for the individual and simultaneous detection of two high-risk human papillomavirus DNA sequences, HPV16E7p and HPV45E6 that exhibits high sensitivity and selectivity is reported. The optimum conditions for surface chemistry preparation and detection of hybridised target were investigated. The LOD obtained are in the pM range, which are sufficient for most real RNA/DNA samples obtained from PCR amplification, usually in the nanomolar range. In a multiplexed detection format, high selectivity was observed over the non-specific sequence, opening the way for the development of an electrochemical high throughput screening assay for multiple high-risk DNA sequences.
Assuntos
Técnicas Biossensoriais/instrumentação , Genes Virais , Papillomaviridae/genética , Sequência de Bases , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/estatística & dados numéricos , Sondas de DNA de HPV/genética , DNA Viral/análise , DNA Viral/genética , Técnicas Eletroquímicas , Papillomavirus Humano 16/genética , Humanos , Análise de Sequência com Séries de Oligonucleotídeos/instrumentação , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Análise de Sequência com Séries de Oligonucleotídeos/estatística & dados numéricos , Proteínas Oncogênicas Virais/genética , Papillomaviridae/classificação , Proteínas E7 de Papillomavirus/genéticaRESUMO
OBJECTIVES: To investigate the prevalence rates of specific human papillomavirus (HPV) types infecting women in Liaoning Province, China. METHODS: Specimens from 4780 patients with cervical disease and 165 age-matched controls were tested for HPV genotypes using a chip hybridization assay. RESULTS: The infection rates were 35.66% for patients with cervicitis, 54.61% for those with ASCUS, 64.14% for those with CIN, 83.76% for those with cervical cancer in situ, and 83.12% for those with invasive cervical cancer. The most common HPV genotype was HPV-16, followed by HPV-58, HPV-52, HPV-33, HPV-53, and HPV-31. There were 1529 single and 731 multiple infections among the 4780 patients. Single infections with high-risk genotypes were associated with various cervical diseases. HPV-16 was present in 399 of the patients with multiple infections. CONCLUSION: Compared with prevalence rates for other populations, the rates of specific HPV types infecting women are different in Liaoning Province of China.
Assuntos
Papillomaviridae/classificação , Infecções por Papillomavirus/virologia , Displasia do Colo do Útero/virologia , Neoplasias Uterinas/virologia , Saúde da Mulher , Adolescente , Adulto , China/epidemiologia , Sondas de DNA de HPV/genética , Feminino , Papillomavirus Humano 16/classificação , Papillomavirus Humano 16/genética , Papillomavirus Humano 16/isolamento & purificação , Humanos , Pessoa de Meia-Idade , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/epidemiologia , Prevalência , Neoplasias Uterinas/epidemiologia , Adulto Jovem , Displasia do Colo do Útero/epidemiologiaRESUMO
In this study, typing of human papilloma virus (HPV) was performed in squamous cell carcinomas of the lung (n=26) as well as putative primaries of head and neck (n=21) and female genital tract (n=5) of the same patients, to test whether additional information to discriminate lung primaries from metastases can be gained by a direct comparison of the HPV status in both tumors. In 3 (14.2%) patients with head and neck as well as lung squamous cell carcinoma, an identical HPV subtype could be detected in both tumors suggesting metastatic disease. In 9 (42.9%) cases, discordant HPV status strongly suggested secondary primaries of the lung. In the remaining 9 (42.9%) patients, no HPV was evident in either tumor. In all patients with carcinomas of the cervix uteri an identical HPV subtype was detected in the cervical and in the lung tumor. In conclusion, the results suggest HPV typing, a method routinely used in cervical biopsies for years, as a very useful diagnostic tool to discriminate primary from metastatic squamous cell carcinoma of the lung, which in our cohort in 57.1% of cases allowed for almost definite classification.
Assuntos
Carcinoma de Células Escamosas/diagnóstico , Sondas de DNA de HPV/genética , Neoplasias de Cabeça e Pescoço/diagnóstico , Neoplasias Pulmonares/diagnóstico , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Neoplasias do Colo do Útero/diagnóstico , Adulto , Idoso , Carcinoma de Células Escamosas/virologia , DNA Viral/análise , Diagnóstico Diferencial , Feminino , Neoplasias de Cabeça e Pescoço/virologia , Humanos , Neoplasias Pulmonares/virologia , Masculino , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular , Metástase Neoplásica/diagnóstico , Análise de Sequência com Séries de Oligonucleotídeos , Papillomaviridae/genética , Infecções por Papillomavirus/virologia , Neoplasias do Colo do Útero/virologiaRESUMO
Neuroendocrine tumours are the second most common laryngeal neoplasms, following squamous carcinoma. In this paper, we report the case of a moderately differentiated neuroendocrine carcinoma NEC (atypical carcinoid) of the larynx in a heavy smoker 67-year-old woman, with a history of hoarseness, dysphagia and dyspnea. The lesion was biopsied and microscopic examination revealed moderately differentiated NEC; thus the patient underwent supraglottic laryngectomy with lymphadenectomy. Here, we emphasized the morphological criteria for a correct pathological diagnosis. Moreover, because it has been demonstrated that many neuroendocrine neoplasms and malignant lesions of the larynx can be related to human papilloma virus (HPV), for the first time, we probed to verify if laryngeal neuroendocrine carcinoma could be due to an HPV infection by using polymerase chain reaction amplification (PCR) of tumoural DNA. On immunohistochemical analysis, the lesion characteristically revealed both neuroendocrine and epithelial differentiation with diffuse staining for chromogranin A, synaptophysin and neuron-specific enolase (NSE) and epithelial membrane antigen (EMA) and overexpression of p53 protein. PCR of NEC DNA did not show any signal for HPV DNA. Thus, this neoplasm is not due to an HPV infection, but a mutation of p53 gene which could cause immunohistochemical overexpression of p53 protein.
Assuntos
Tumor Carcinoide/patologia , Neoplasias Laríngeas/patologia , Idoso , Biomarcadores Tumorais/análise , Biópsia , Tumor Carcinoide/genética , Tumor Carcinoide/cirurgia , Análise Mutacional de DNA , Sondas de DNA de HPV/genética , DNA de Neoplasias/genética , Feminino , Humanos , Neoplasias Laríngeas/genética , Neoplasias Laríngeas/cirurgia , Laringectomia , Laringe/patologia , Excisão de Linfonodo , Invasividade Neoplásica , Estadiamento de Neoplasias , Reação em Cadeia da Polimerase , Proteína Supressora de Tumor p53/genéticaRESUMO
Squamous cell carcinoma (SCC) in larynx is rare with children and adolescents. Usually larynx cancer is common with male smokers in the 7th decade. Among patients with no history of tobacco and/or alcohol consumption several factors have can play a role in the outbreak of laryngeal cancer: such as individual predisposition, radiation, gastroesophageal reflux, viral infection, dietary factors and environmental influences. In literature only few cases of laryngeal cancer with children are reported. Recent studies show that the most frequent laryngeal malignancy is the embryonal rhabdomyosarcoma. Besides the recurrent respiratory papillomatosis (RRP) based on an infection with human papilloma virus (HPV) types 6 and 11 (low risk) and types 16 and 18 (high risk) is known for a possible malignant transformation towards a SCC. HPV type 26 is only reported as low risk type HPV associated with cervical cancer. Final diagnosis often takes a long time. Initial symptoms such as hoarseness, cough or shortness of breath are often referred to more typical pediatric diseases or laryngeal development.