Universal human papillomavirus typing by whole genome sequencing following target enrichment: evaluation of assay reproducibility and limit of detection.

BACKGROUND: We recently described a method for unbiased detection of all known human papillomaviruses (HPV) types with the potential for the determination of their variant and integration from the resulting whole genome sequence data. Considering the complex workflow for target-enriched next generation sequencing (NGS), we focused on the reproducibility and limit of detection (LOD) of this new universal HPV typing assay in this study. RESULTS: We evaluated the reproducibility and LOD for HPV genotyping based on our recently published method that used RNA-baits targeting whole genomes of 191 HPV types, Agilent SureSelect protocol for target enrichment and Illumina HiSeq 2500 for sequencing (eWGS, enriched whole genome sequencing). Two libraries, prepared from pooled plasmids representing 9 vaccine HPV types at varying input (1-625 copies/reaction), were sequenced twice giving four replicates for evaluating reproducibility and LOD. eWGS showed high correlation in the number of reads mapped to HPV reference genomes between the two flow-cell lanes within (R2 = 1) and between experiments (R2 = 0.99). The number of mapped reads was positively correlated to copy number (ß = 13.9, p < 0.0001). The limit of blank (LOB) could be calculated based on mapped reads to HPV types not included in each sample. HPV genotyping was reproducible for all 9 types at 625 copies using multiple cut-off criteria but LOD was 25 copies based on number of reads above LOB even when multiple types were present. eWGS showed no bias for HPV genotyping under single or multiple infection (p = 0.16-0.99). CONCLUSIONS: The universal eWGS method for HPV genotyping has sensitivity, competitive with widely used consensus PCR methods with reduced type competition, and with the potential for determination of variant and integration status. The protocol used in this study, using defined samples varying in complexity and copy number, analyzed in replicate and duplicate assays, is applicable to most WGS methods.

Molecular progression to cervical precancer, epigenetic switch or sequential model?

The evolution of precancerous cervical lesions is poorly understood. A widely held model of cervical intraepithelial neoplasia grade 3 (CIN3) development is sequential progression from normal through CIN1 and CIN2 to CIN3. Another hypothesis, the "molecular switch" model, postulates that CIN3 can evolve directly from human papillomavirus (HPV)-infected normal epithelium without progressing through CIN1 and CIN2. To shed light on this process, we compared DNA methylation of selected human biomarkers and HPV types in two groups of CIN1: CIN1 that were near or adjacent to CIN3 (adjacent-CIN1) and CIN1 that were the principal lesions with no CIN3 detected (principal-CIN1). 354 CIN (CIN1 and CIN3) and normal tissue areas were dissected and typed for HPV from 127 women who underwent loop electrosurgical excision procedures (LEEP). Methylation of genes EPB41L3 and the viral regions of HPV16-L1/L2, HPV18-L2, HPV31-L1, and HPV33-L2 were determined by a highly accurate quantitative pyrosequencing of bisulfite converted DNA. There was a significant trend of increased methylation with disease grade comparing normal to CIN1 and CIN3 (p < 0.0001). Adjacent-CIN1 predominantly shared the same HPV types as the CIN3, however, methylation differed substantially between adjacent-CIN1 and CIN3 (p = 0.008). In contrast diagnostically principal-CIN1 had an indistinguishable methylation distribution compared to adjacent-CIN1 (EPB41L3: p = 0.49; HPVme-All: p = 0.11). Our results suggest that progression from normal epithelium to CIN1 or CIN3 is usually promoted by the same HPV type but occurs via distinct DNA epigenotypes, thus favoring the "molecular switch" model.

Universal Human Papillomavirus Typing Assay: Whole-Genome Sequencing following Target Enrichment.

We designed a universal human papillomavirus (HPV) typing assay based on target enrichment and whole-genome sequencing (eWGS). The RNA bait included 23,941 probes targeting 191 HPV types and 12 probes targeting beta-globin as a control. We used the Agilent SureSelect XT2 protocol for library preparation, Illumina HiSeq 2500 for sequencing, and CLC Genomics Workbench for sequence analysis. Mapping stringency for type assignment was determined based on 8 (6 HPV-positive and 2 HPV-negative) control samples. Using the optimal mapping conditions, types were assigned to 24 blinded samples. eWGS results were 100% concordant with Linear Array (LA) genotyping results for 9 plasmid samples and fully or partially concordant for 9 of the 15 cervical-vaginal samples, with 95.83% overall type-specific concordance for LA genotyping. eWGS identified 7 HPV types not included in the LA genotyping. Since this method does not involve degenerate primers targeting HPV genomic regions, PCR bias in genotype detection is minimized. With further refinements aimed at reducing cost and increasing throughput, this first application of eWGS for universal HPV typing could be a useful method to elucidate HPV epidemiology.

[Human papillomavirus associated cervix uteri morbidity in Hungary: epidemiology and correlation with the HPV types and the simultaneous cytological diagnosis]. / Humán papillomavírus asszociált méhnyak-megbetegedések Magyarországon: epidemiológia és a HPV-típusok összefüggése a párhuzamosan végzett citológiai diagnózissal.

INTRODUCTION: Persistent infection of human papillomavirus is known to cause cervical intraepithelial neoplasia or cancer in the cervix uteri and other HPV-associated cancers in different localization. Based on epidemiological and biological data, principally the high risk HPV is responsible for development of cervical these cancers. However, we have no information about the frequently distribution of different HPV types and what is the correlation between the HPV types and cytological diagnosis in cervical intraepithelial neoplasia (CIN). AIM: In this paper, we are going to present new data involving incidence and mortality of HPV-associated cancers during the period of 2009-2015 in Hungary. We are also going to investigate the correlation of cervical cytological diagnosis and HPV typing, and the preventive effect of HPV vaccination. METHOD: The epidemiological data spring from the National Cancer Registry. HPV typing was performed by Linear Array HPV Genotyping Test. Simultaneous cytological diagnosis and HPV typing was carried out on 2048 cytological samples collected in period of 2009-2016. RESULTS: According to the epidemiologic data, the most frequently occurring HPV-associated cancer is the laryngeal carcinoma in man, and the cervical cancer in woman in Hungary. During the 2009-2015 time intervals, the frequency distribution of head and neck cancers was not changed in man, but the incidence of tongue root squamous cell carcinomas was gradually increasing in woman. We have defined the clinical significance of single and simultaneously multiple HPV infection and have investigated the correlation of the HPV frequency distribution and cytological diagnosis in CIN. It was found that in the cytological negativity of probably/possibly carcinogen pHR-HPV group classified by IACR was much more frequent as in HR-HPV group (56% versus 47%). The presence of simultaneous multiplex HPV infection betokens an increased cancer risk. According to the international publications, the ratio of HPV16 just twice as big as in cervical cancer, what we found in CIN (60% versus 30%). The frequency order of the HPV18 is 2nd in cancer, and 9th in CIN. Comparing the frequency distribution of HR/pHR-HPVs in cervical cancer and CIN, the HR-HPV35 is very rarely occurring in CIN, the pHR-HPV56, 66, and 73 is more frequently seen in CIN as in carcinoma. Appreciated the preventive value of anti-HPV vaccines, we have found a significant differences in group with 1 HPV/sample and in group with more than 1 HPV/sample. CONCLUSION: The frequency distribution of tongue root squamous cell carcinoma and cervical cancer was gradually increasing in woman. The overall preventive effect of 9-valent vaccine is 80.3%. This preventive value should be higher because of the transformation ability of the different HPV types is not same. Out of consideration for HPV incidence in cancer, the preventive effect of 9-valent or 4-valent vaccines might reach to 93% or 73%. However, the pHR-HPVs are biologically active, it is not sufficient for the inclusion of these HPV types into population-wide HPV-DNA based cervical screening programs. Orv Hetil. 2017; 158(31): 1213-1221.

Human papillomavirus prevalence in oropharyngeal cancer before vaccine introduction, United States.

We conducted a study to determine prevalence of HPV types in oropharyngeal cancers in the United States and establish a prevaccine baseline for monitoring the impact of vaccination. HPV DNA was extracted from tumor tissue samples from patients in whom cancer was diagnosed during 1995-2005. The samples were obtained from cancer registries and Residual Tissue Repository Program sites in the United States. HPV was detected and typed by using PCR reverse line blot assays. Among 557 invasive oropharyngeal squamous cell carcinomas, 72% were positive for HPV and 62% for vaccine types HPV16 or 18. Prevalence of HPV-16/18 was lower in women (53%) than in men (66%), and lower in non-Hispanic Black patients (31%) than in other racial/ethnic groups (68%-80%). Results indicate that vaccines could prevent most oropharyngeal cancers in the United States, but their effect may vary by demographic variables.

Extended HPV typing test performed better predict value for CIN2+ among elderly women in China.

Objective: The aim of this study was to examine the cervical cancer screening practices among women residing in Lingang New District of Shanghai. Moreover, the study aimed to delve into the characteristics of HPV infection and cervical lesions in older women (≥60 years old), seeking for more effective method for cervical cancer screening. Methods: This is a cross-sectional study enrolled women who were referred to colposcopy and cervical histological examination due to abnormal cytology or HPV tests from Shanghai Sixth People's Hospital between January 2018 and December 2022. Results: A total of 1,931 women (mean age: 41.8 ± 12.5, range: 18-88 years old) were enrolled, 119 individuals aged ≥ 60 and 1732 aged <60. The infection rates of HPV52, 33, 35, 56, 26 and 81 were significantly higher in the elderly group. Multiple HPV infection rates were also higher in this group and were associated with cervical lesions. The probability of LSIL, HSIL and Ca in women over 60 years old was significantly higher compared to women under 60. The top three HPV genotypes in elderly women with CIN2+ were HPV16, 52, and 58. The Yoden index was higher for extended typing for HPV 31/33/45/52/58(0.41) compared to cytology(0.29), high risk HPV without specific typing(0.07), cotest(cytology and high risk HPV, 0.06 or 0.30), or the current shunt strategy(0.07). Conclusions: Elderly women still need to continue cervical cancer screening, and extended typing test for HPV16/18/31/33/45/52/58 is a more effective method for this age group.

Validation in Zambia of a cervical screening strategy including HPV genotyping and artificial intelligence (AI)-based automated visual evaluation.

BACKGROUND: WHO has recommended HPV testing for cervical screening where it is practical and affordable. If used, it is important to both clarify and implement the clinical management of positive results. We estimated the performance in Lusaka, Zambia of a novel screening/triage approach combining HPV typing with visual assessment assisted by a deep-learning approach called automated visual evaluation (AVE). METHODS: In this well-established cervical cancer screening program nested inside public sector primary care health facilities, experienced nurses examined women with high-quality digital cameras; the magnified illuminated images permit inspection of the surface morphology of the cervix and expert telemedicine quality assurance. Emphasizing sensitive criteria to avoid missing precancer/cancer, ~ 25% of women screen positive, reflecting partly the high HIV prevalence. Visual screen-positive women are treated in the same visit by trained nurses using either ablation (~ 60%) or LLETZ excision, or referred for LLETZ or more extensive surgery as needed. We added research elements (which did not influence clinical care) including collection of HPV specimens for testing and typing with BD Onclarity™ with a five channel output (HPV16, HPV18/45, HPV31/33/52/58, HPV35/39/51/56/59/66/68, human DNA control), and collection of triplicate cervical images with a Samsung Galaxy J8 smartphone camera™ that were analyzed using AVE, an AI-based algorithm pre-trained on a large NCI cervical image archive. The four HPV groups and three AVE classes were crossed to create a 12-level risk scale, ranking participants in order of predicted risk of precancer. We evaluated the risk scale and assessed how well it predicted the observed diagnosis of precancer/cancer. RESULTS: HPV type, AVE classification, and the 12-level risk scale all were strongly associated with degree of histologic outcome. The AVE classification showed good reproducibility between replicates, and added finer predictive accuracy to each HPV type group. Women living with HIV had higher prevalence of precancer/cancer; the HPV-AVE risk categories strongly predicted diagnostic findings in these women as well. CONCLUSIONS: These results support the theoretical efficacy of HPV-AVE-based risk estimation for cervical screening. If HPV testing can be made affordable, cost-effective and point of care, this risk-based approach could be one management option for HPV-positive women.

A human papillomavirus whole genome plasmid repository: A resource for HPV DNA quality control reagents.

Well characterized reference reagents are useful for assay validation, proficiency/competency assessment, daily run controls, and to improve inter-laboratory comparisons. Synthetic human papillomavirus (HPV) DNA fragments and plasmid clones are available, but synthetic fragments include limited segments of the HPV genome and many HPV plasmids have interrupted coding regions or contain partial genomes. As a result, they are not compatible with all HPV DNA detection and typing assays. To address this need, we are establishing an HPV plasmid repository of HPV clones containing the whole genome of each type with no interruptions in coding regions. To date, HPV plasmid clones for 16 HPV types, (including all vaccine types and 14 types in clinical assays: HPV6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68) have been constructed using a Gibson assembly method and validated by sequencing and the Novaplex HPV typing assay. The newly constructed HPV whole genome plasmids can serve as a quality control reagent resource for HPV DNA assays and are available for public health and research laboratories.

Broad-Spectrum Detection of HPV in Male Genital Samples Using Target-Enriched Whole-Genome Sequencing.

Most human papillomavirus (HPV) surveillance studies target 30-50 of the more than 200 known types. We applied our recently described enriched whole-genome sequencing (eWGS) assay to demonstrate the impact of detecting all known and novel HPV types in male genital samples (n = 50). HPV was detected in nearly all (82%) samples, (mean number of types/samples 13.6; range 1-85), and nearly all HPV-positive samples included types in multiple genera (88%). A total of 560 HPV detections (237 unique HPV types: 46 alpha, 55 beta, 135 gamma, and 1 mu types) were made. The most frequently detected HPV types were alpha (HPV90, 43, and 74), beta (HPV115, 195, and 120), and gamma (HPV134, mSD2, and HPV50). High-risk alpha types (HPV16, 18, 31, 39, 52, and 58) were not common. A novel gamma type was identified (now officially HPV229) along with 90 unclassified types. This pilot study demonstrates the utility of the eWGS assay for broad-spectrum type detection and suggests a significantly higher type diversity in males compared to females that warrants further study.

Multiple-type HPV infection predicting condyloma acuminatum recurrence after aminolevulinic acid photodynamic therapy.

BACKGROUND: Aminolevulinic acid photodynamic therapy (ALA-PDT) has been widely used in the treatment of condyloma acuminatum (CA), but there is a lack of reports on clinical factors related to CA relapse after photodynamic therapy. METHODS: The clinical data of patients with CA treated with ALA-PDT from April 2018 to December 2019 were retrospectively analyzed, including HPV type, follow-up time and recurrence within 6 months after treatment. The patients were classified into single-type HPV infection and multiple-type HPV infection groups. Besides, the patients were also classified into high-risk HPV infection, low-risk HPV infection, and low + high-risk HPV infection groups. Univariate and multivariate COX regression was performed to analyze whether HPV type was related with CA relapse after photodynamic therapy. RESULTS: A total of 161 CA patients who underwent ALA-PDT were analyzed in this study. CA recurred in 20 patients within 6 months after treatment, with a recurrence rate of 12.4%. Of them, the patients with multiple-type HPV infection made up 85%. Multivariate COX regression analysis revealed that multiple-type HPV infection was associated with CA recurrence (HR:5.0; 95% CI: 1.1-21.4; P = 0.032). Of the patients with CA recurrence, 70% developed low + high-risk infections, a proportion significantly higher than that in patients without CA relapse (16.3%, P < 0.001). Using low-risk infection as a reference, low + high-risk infection was positively correlated with postoperative recurrence (HR: 6.7; 95% CI: 1.6-26.2; P = 0.006). CONCLUSION: Multiple-type HPV infection were closely associated with CA recurrence after photodynamic therapy.

Implementation of the Program for Early Detection of Cervical Cancer in the Federal Republic of Germany.

INTRODUCTION: There are many cervical cancer screening programs in the world. Germany, as well as a number of other countries in the world, carry out its national cervical cancer screening program. AIM: In order to improve screening results, new guidelines for cervical cancer screening are in force in Germany. METHODS: Auhors used descriptive-analytical method to described advantages and disadvantages of new adopted Guidelines for screnning program according eperiences of used previous program in Germany. DISCUSSION: These guidelines have been adopted and approved by the competent Federal Committee for the implementation of cervical cancer screening in Germany. The committee is under the independent management of doctors and health insurance companies. The Committee is also under the legal control of the German Federal Ministry of Health. CONCLUSION: New Guidelines for Cervical Cancer Screening in Germany has an unchanged part relating to cervical cytodiagnostics. In addition, HPV typizatiion has been integrated in the new screening guidelines to further improve the quality of cervical cancer screening in Germany.

Multiple HPV Infections and Viral Load Association in Persistent Cervical Lesions in Mexican Women.

Persistent high-risk human papillomavirus (HR-HPV) infections play a major role in the development of invasive cervical cancer (CC), and screening for such infections is in many countries the primary method of detecting and preventing CC. HPV typing can be used for triage and risk stratification of women with atypical squamous cells of undetermined significance (ASC-US)/low-grade cervical lesions (LSIL), though the current clinical practice in Mexico is to diagnose CC or its preceding conditions mainly via histology and HR-HPV detection. Additional information regarding these HPV infections, such as viral load and co-infecting agents, might also be useful for diagnosing, predicting, and evaluating the possible consequences of the infection and of its prevention by vaccination. The goal of this follow-up hospital case study was to determine if HPV types, multiple HPV infections, and viral loads were associated with infection persistence and the cervical lesion grade. A total of 294 cervical cytology samples drawn from patients with gynecological alterations were used in this study. HPV types were identified by real-time PCR DNA analysis. A subset of HPV-positive patients was reevaluated to identify persistent infections. We identified HPV types 16, 18, and 39 as the most prevalent. One hundred five of the patients (59%) were infected with more than one type of HPV. The types of HPV associated with multiple HPV infections were 16, 18, and 39. In the follow-up samples, 38% of patients had not cleared the initially detected HPV infection, and these were considered persistent. We found here an association between multiple HPV infections and high viral loads with and infection persistence. Our findings suggest there are benefits in ascertaining viral load and multiple HPV infections status of HR-HPV infections for predicting the risk of persistence, a requirement for developing CC. These findings contribute to our understanding of HPV epidemiology and may allow screening programs to better assess the cancer-developing risks associated with individual HR-HPV infections.

Bioinformatics Pipeline for Human Papillomavirus Short Read Genomic Sequences Classification Using Support Vector Machine.

We recently developed a test based on the Agilent SureSelect target enrichment system capturing genomic fragments from 191 human papillomaviruses (HPV) types for Illumina sequencing. This enriched whole genome sequencing (eWGS) assay provides an approach to identify all HPV types in a sample. Here we present a machine learning algorithm that calls HPV types based on the eWGS output. The algorithm based on the support vector machine (SVM) technique was trained on eWGS data from 122 control samples with known HPV types. The new algorithm demonstrated good performance in HPV type detection for designed samples with 25 or greater HPV plasmid copies per sample. We compared the results of HPV typing made by the new algorithm for 261 residual epidemiologic samples with the results of the typing delivered by the standard HPV Linear Array (LA). The agreement between methods (97.4%) was substantial (kappa= 0.783). However, the new algorithm identified additionally 428 instances of HPV types not detectable by the LA assay by design. Overall, we have demonstrated that the bioinformatics pipeline is an accurate tool for calling HPV types by analyzing data generated by eWGS processing of DNA fragments extracted from control and epidemiological samples.

Comparison of the novel human papillomavirus 4 auto-capillary electrophoresis test with the hybrid capture 2 assay and with the PCR HPV typing set test in the detection of high-risk HPV including HPV 16 and 18 genotypes in cervical specimens.

The aim of this study was to compare the novel human papillomavirus (HPV) detection method, the HPV 4 Auto-capillary Electrophoresis (ACE) test with the hybrid capture (HC) 2 assay for the detection of high-risk HPVs. In addition, we compared the HPV 4 ACE test with the polymerase chain reaction HPV Typing Set test for the detection of HPV 16 and HPV 18 genotypes. One hundred ninety-nine cervical swab samples obtained from women with previous abnormal Pap smears were subjected to testing with the three HPV tests. The HPV 4 ACE test and the HC 2 assay showed substantial agreement for detection of high-risk HPVs (85.4%, kappa=0.71). The HPV 4 ACE test also showed substantial agreement with the PCR HPV Typing Set test in the detection of HPV 16 and HP V 18 genotypes (89.9%, kappa=0.65). In correlation with cytologic results, the sensitivities and specificities of the HPV 4 ACE test and HC 2 assay were 92.9% vs. 92.9% and 48.1% vs. 50.8%, respectively, when high-grade squamous intraepithelial lesions were regarded as abnormal cytologies. The novel HPV 4 ACE test is a valuable tool for the detection of high-risk HPVs and for genotyping of HPV 16 and HPV 18.

Bowenoid papulosis successfully treated with imiquimod 5% cream.

A 24-year-old healthy Japanese female was diagnosed as having bowenoid papulosis in the genital area. The histopathological findings revealed acanthosis, papillomatosis, dyskeratotic cells and clumping cells with mild atypical nuclei. Human papillomavirus type 16 was detected in the lesion. The lesion was successfully treated with topical imiquimod 5% cream after 2 months. Imiquimod 5% cream is a potentially effective treatment modality for lesions that are difficult to treat with surgical excision.

Potential opportunities to reduce cervical cancer by addressing risk factors other than HPV.

Cervical cancer is the most common cancer in developing world and 80% of global burden is reported from these nations. Human papillomavirus along with poverty, illiteracy/lower education level and standards, multi-parity, tobacco, malnutrition and poor genital hygiene may act synergistically to cause cervical cancer. Risk factor of cervical cancer may in itself be the reason for non-viability of cervical cancer vaccine program in this part of the world. Interventions to address these risk factors in addition to vaccination of girls before their sexual debut may hold promises of reducing the morbidity and mortality of female genital cancers.

Detección y tipificación de virus del papiloma humano en biopsias de carcinoma ductal infiltrante y lesiones benignas de mama en mujeres venezolanas

Objetivo: Realizar la detección y tipificación del virus del papiloma humano (VPH) en muestras de biopsias de tejido mamario con carcinoma ductal infiltrante. Métodos: Estudio descriptivo de corte transversal de 57 biopsias de carcinoma ductal infiltrante, y 41 biopsias de lesiones benignas de mama de pacientes venezolanas, estas fueron evaluadas utilizando la técnica PCR-RFLP en busca de la presencia del genoma del virus de papiloma humano. El riesgo OR fue evaluado mediante análisis estadístico con el paquete SPSS 12.0. Resultados: Treinta y tres (57,9 %) de las muestras de carcinoma ductal infiltrante tuvieron un resultado positivo para virus de papiloma humano, 19 de ellas pudieron ser tipificadas como: VPH-6b 15,15 %; VPH-11 3,03 %; VPH-18 12,12 %; VPH-33 27,27 %; VPH-45 3,03 % y VPH-58 3,03 %; de este grupo el 42,4 % fueron positivas no determinadas para la presencia de ADN del virus. Seis biopsias de lesiones benignas (14,6 %), presentaron infección por virus de papiloma humano, determinándose para ellas los tipos VPH-6b 33,33 %, VPH-11 16,67%, VPH-33 16,67% y 33,33 % positivas no determinadas. Se determinó estadísticamente que la presencia de virus de papiloma humano en tejido mamario aumenta 10,77 veces la posibilidad de desarrollar carcinoma ductal infiltrante. Conclusiones: Los hallazgos corroboran los resultados de otros investigadores, colocando al virus de papiloma humano como posible agente involucrado en la inmortalización de las células epiteliales de la mama.

Objective: To perform the detection and typing of human papilloma (HPV) virus in biopsy samples of breast tissue invasive ductal cancer. Methods: Cross-sectional study of 57 biopsies of invasive ductal carcinoma, and 41 biopsies of benign breast lesions of Venezuelan patients were evaluated using the PCR-RFLP technique for the presence of the human papillomavirus genome. The OR risk was evaluated by statistical analysis using SPSS package. Results: Thirty-three (57.9%) of invasive ductal carcinoma samples had a positive result for human papillomavirus, 19 of them could be classified as: HPV-6b 15.15%; HPV-11 3.03%; HPV-18 12.12%; HPV-33 27.27%; HPV-45 3.03% and HPV-58 3.03%. This group 42.4% were positive not determined for the presence of virus DNA. Six biopsies of benign lesions (14.6%) had human papillomavirus infection, determining for themselves the types HPV-6b 33.33%, 16.67% HPV-11, HPV-33 16.67% and 33.33% not determined positive. It is statistically determined that the presence of human papillomavirus in breast tissue 10.77 times increases the possibility of developing invasive ductal carcinoma. Conclusions: These findings corroborate the results of other researchers, placing human papillomavirus as a possible agent involved in the immortalization of epithelial cells of the breast.

Prevalence of human papillomavirus infection, distribution of viral types and risk factors in cervical samples from human immunodeficiency virus-positive women attending three human immunodeficiency virus-acquired immune deficiency syndrome reference centres in northeastern Brazil

Human immunodeficiency virus (HIV)-positive patients have a greater prevalence of coinfection with human papillomavirus (HPV) is of high oncogenic risk. Indeed, the presence of the virus favours intraepithelial squamous cell lesion progression and may induce cancer. The aim of this study was to evaluate the prevalence of HPV infection, distribution of HPV types and risk factors among HIV-positive patients. Cervical samples from 450 HIV-positive patients were analysed with regard to oncotic cytology, colposcopy and HPV presence and type by means of polymerase chain reaction and sequencing. The results were analysed by comparing demographic data and data relating to HPV and HIV infection. The prevalence of HPV was 47.5%. Among the HPV-positive samples, 59% included viral types of high oncogenic risk. Multivariate analysis showed an association between HPV infection and the presence of cytological alterations (p = 0.003), age greater than or equal to 35 years (p = 0.002), number of partners greater than three (p = 0.002), CD4+ lymphocyte count < 200/mm3 (p = 0.041) and alcohol abuse (p = 0.004). Although high-risk HPV was present in the majority of the lesions studied, the low frequency of HPV 16 (3.3%), low occurrence of cervical lesions and preserved immunological state in most of the HIV-positive patients were factors that may explain the low occurrence of precancerous cervical lesions in this population.

Tipificación del virus del papiloma humano en muestras cervicales de 15 mujeres atendidas en el Instituto Nacional del Cáncer, Año 2007 / Typing of human papilloma virus in cervical samples of fifteen women that attended the National Institute of Cancer in December, 2007

El cáncer de cuello uterino es la segunda neoplasia maligna en la población femenina anivel mundial siendo el virus de papiloma humano (HPV) la causa principal. El objetivo deeste estudio preliminar fue determinar el tipo de HPV en mujeres atendidas en el InstitutoNacional del Cáncer en Diciembre del 2007, por amplificación de cadena de la polimerasaasociado a digestión por enzimas de restricción (PCR-RFLP) y observar la frecuencia deotros factores de riesgo asociados al cáncer. HPV fue detectado en 14 de 15 mujeres conresultados anormales de citología y/o colposcopía. Ocho de las 14 mujeres fueronpositivas para HPV de alto riesgo – HR HPV (tipos 16, 31, 58, 33, 45), presentando 6 delas 8 mujeres resultados de biopsia de neoplasia intraepitelial cervical I y III (CIN I y CINIII). Se observó un caso de infección múltiple con los tipos de HR-HPV 33 y 45. Fueronobservados 7 casos con falta de concordancia entre los resultados de citología,colposcopía y biopsia, en los cuales la detección de tipos de HR-HPV contribuyó aidentificar las mujeres con mayor riesgo de desarrollar CIN. Se observó en algunos casosla presencia de otros factores de riesgo para CIN, como el consumo de cigarrillo por 10 y30 años y el uso de anticonceptivos orales por 20 años. En conclusión, los resultadospreliminares sugieren que la detección de tipos de HPV por PCR-RFLP puede ser útil paraorientar el manejo del paciente.