Cervical Brush Research Articles

Hormonal regulation of oviductal glycoprotein 1 (OVGP1; MUC9) in the macaque cervix: a novel indicator of progestogen action

Progestogen-only contraception is reported to act by suppressing ovulation and/or altering cervical mucus secretion. Unfortunately, the effect of contraceptives on mucus quality is difficult to assess accurately by traditional Insler score or post-coital tests, and the lack of a molecular test for mucus quality has created a roadblock to the development of new contraceptives that target the cervix. Oviductal glycoprotein 1 (OVGP1; MUC9) is a hormonally regulated, carbohydrate-rich glycoprotein secreted by oviductal epithelial cells [1]. In addition to the oviduct, the OVGP1 expression has been reported for the cervix of rabbits, [2] mice and women, but hormonal regulation of OVGP1 in the cervix is poorly understood. The objective of this study was to characterize hormonal regulation and localization of OVGP1 in the macaque cervix as a novel marker of progestogen action. Real-time PCR and immunohistochemical analysis of the macaque cervix. Adult ovariectomized rhesus macaques (Macaca mulatta) were treated sequentially with estradiol (E2) and then E2 plus progesterone (P) to create artificial menstrual cycles. Reproductive tracts were collected in the artificial proliferative phase (E2 alone; n=13) and in the artificial secretory phase after 1, 3, 7, and 14 days of P (n=4, 5, 5, and 10, respectively). Total RNA was isolated from cervical samples and analyzed for expression of OVGP1 by TaqMan® real-time PCR and presented relative to expression of ribosomal (S10) RNA. Representative samples were analyzed by immunohistochemistry on cryosections with polyclonal antibodies specific to OVGP1. Representative sections were also stained for estrogen receptor 1 (ESR1) and progesterone receptor (PGR) as independent assays for E2 and (P) action. OVGP1 transcript was upregulated in the proliferative phase, maximal in the late proliferative phase, and maintained through day 1 of the secretory phase. There was a significant decrease (P<0.01) in OVGP1 expression between secretory phase day 1 and day 3 and a further significant (P<0.05) decrease by secretory phase day 7 and 14. IHC revealed specific OVGP1 staining in columnar cells of the mid cervix. Staining was strongest in the apical portion of the epithelial cells consistent with secretion into the cervix mucus. As expected, ESR-1 and PGR was present in the cervix throughout the cycle. ESR1 and PGR staining was stronger in the cervix epithelium during the proliferative phase of the cycle. These results indicate that OVGP1 is expressed in the primate cervix as well as the primate oviduct, and strongly upregulated by estrogen in the proliferative phase and down-regulated by P during the secretory phase. The cervical epithelium would be easily accessible by biopsy or cervical brush, and OVGP1 could be assayed either by ELISA or by real-time PCR to provide a rapid and very sensitive method for identifying the effect of progestogen-based contraceptives on the cervix in clinical trials.

Abstract LB-133: Triage of high risk HPV positive women before colposcopy with reflex tests in pap smears and screening of high risk HPV in Transrenal DNA isolated from urine, using novel workflows to identify panels of methylated viral and human DNA

Abstract We set out to develp novel workflows to identify panels of methylated human papilloma virus (HPV) and human genes that can discriminate between CIN2+ and normal/CIN1 patients in liquid prep samples and in Transrenal DNA (TrDNA) isolated from urine. Using Human DNA methylation arrays and massively parallel Next Generation Sequencing (NGS) for Discovery and quantitative Methylation Specific PCR (qMSP) for validation, we found that promoter methylation of ZNF516, FKBP6, and INST1 discriminates samples with CIN2+ lesions from no intraepithelial lesions or malignancy (NILM) in cervical brush samples: 88.3% sensitivity, 88.9% specificity, 93.2 Area Under the Curve (AUC), 86.9% positive predictive value (PPV) and 90.2% negative predictive value (NPV). Using custom sequence capture pools of baits, we pulled down genomic and bisulfite converted high-risk HPV DNA before library prep for massively parallel Next Generation Sequencing (NGS) in 454 and MiSeq instruments, respectively to identify high risk HPV and the HPV methylome. Using our NGS results, we designed Syber Greeen qPCR assays to detect high risk HPV DNA in tissue and in Trans Renal DNA (TrDNA). We also designed qMSP primers-probe sets to detect methylated DNA in the HPV16-L1 gene region and ZNF516, FKBP6, and INST1 in TrDNA. TrDNA is circulating DNA that gets filtered by the kidneys and we isolate from urine as fragmented DNA ranging is size from 50 to 250 bp. We replicated the cervical brush epithelium results in liquid prep samples from another cohort, by adding HPV16-L1 methylation to the panel of methylated host genes (ZNF516, FKBP6, and INST1): 90.9% sensitivity, 60.9% specificity, AUC = 90.1, 52.6% positive predictive value (PPV) and 93.3% NPV. These results were verified in plasma DNA (AUC = 80.7) and TrDNA (AUC = 86.1) isolated from a subset of patients who provided liquid prep samples. Our results suggest that a biomarker panel that quantifies DNA methylation in the HPV-LI gene, together with promoter methylation of three host genes, ZNF516, FKBP6, and INST1, may be used as a reflex test in liquid prep to triage high-risk HPV positive women into colposcopy in countries with established cervical cancer screening programs. Our results also suggest that a panel of human and viral DNA methylation markers can be combined with a high-risk HPV DNA assay, to develop urine screening and triage tests in countries where cervical cancer screening programs are not effective due to cultural or infrastructure factors. Country specific DNA methylomes of normal and premalignant cervical epithelium for different age groups can be created as reference methylomes, to better understand the cultural-, socio-economic- and age-specific variations in the cervical epithelium and their correlation with HPV DNA methylomes. Citation Format: Rafael E. Guerrero-Preston, Blanca L. Valle, Anne Jedlicka, Nitesh Turaga, Liliana Florea, Oluwasina Folawiyo, Francesca Pirini, Angelo Vergura, Maartje Noordhuis, Gabriela Perez, Marisa Renehan, Carolina Guerrero-Díaz, Edgardo De Jesus-Rodríguez, Keimari Méndez-Martínez, José Rodríguez-Orengo, Teresa Díaz-Montes, David Sidransky, Josefina Romaguera. Triage of high risk HPV positive women before colposcopy with reflex tests in pap smears and screening of high risk HPV in Transrenal DNA isolated from urine, using novel workflows to identify panels of methylated viral and human DNA. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-133.

Prevalence of high-risk human papilloma virus types and cervical smear abnormalities in female sex workers in Chandigarh, India

Purpose: Cervical cancer is the most common cancer among women in developing nations. Nearly 90% of the cases have been linked to the presence of high-risk human papillomavirus (hrHPV) types 16 and 18. The risk of cervical cancer may be high in female sex workers (FSWs) due to multiple sexual partners. This study aimed to determine the prevalence of cytological abnormalities and hrHPV types 16 and 18 in FSWs in Chandigarh, North India using the liquid-based cytology (LBC) approach. Materials and Methods: The cervical brush samples were collected from 120 FSW and 98 age-matched healthy controls (HCs). These were subjected to pap smear using conventional method, LBC and the detection of hrHPV types 16 and 18 was carried out using polymerase chain reaction. Results: The LBC samples showed better cytological details and also reduced the number of unsatisfactory smears from 11% in Pap to 1.5% in the LBC. A significantly higher number of inflammatory smears were reported in FSWs (51.7% vs. 34.7%, P = 0.01). The hrHPV types 16/18 were detected in 33/120 (27.5%) FSW versus 23/98 (23.5%) HCs. The risk of acquiring hrHPV was higher in FSWs, who had age at first sex ≤25 years, higher income and the habit of smoking. Conclusion: The high prevalence of hrHPV among FSWs and HCs suggests the need for the implementation of effective National Screening Programme for early detection of hrHPV types to decrease the burden of cervical cancer, especially in high-risk population.

Prevalence, Genotype Distribution and Risk Factors for Cervical Human Papillomavirus Infection in the Grand Tunis Region, Tunisia.

Implementation of Human Papillomavirus (HPV) vaccination should be considered a key cervical cancer prevention strategy in Tunisia, where Pap smear screening is not efficient. This study aims to estimate the prevalence and to identify risk factors associated with HPV infection among women from Grand Tunis, Tunisia. We conducted a cross-sectional study, between December 2012 and May 2013. Eligible women for this study were those aged 18–65 years, sexually active, who sought medical attention at their primary health care centre or clinic in Grand Tunis, Tunisia and who gave written consent. A liquid-based Pap smear sample was obtained from all women using a cervical brush. Only women with betaglobin positive test were further analysed for HPV detection and typing. A nested-PCR of the L1 region was performed followed by reverse line blot hybridization to facilitate the specific detection of 31 HPV genotypes. Multiple logistic regression modeling was used for the analysis of associations between variables with some considered possible confounders after checking for interactions. A total of 391 women were enrolled in this study and 325 out of the 391 cervical samples were positive for the betaglobin test. Overall HPV prevalence was 13.2% [9.8%−17.5%], with the following most prevalent HPV genotypes: HPV6 (40%), HPV40 (14%), HPV16 (12%), HPV52 (9%), HPV31 and HPV59 (7%), followed by HPV68 (4%). Mean age of HPV positive women was 40.7±0.92 years. Independently associated risk factors of HPV infection were smoking (OR:2.8 [0.8–9.6]), low income (OR:9.6 [1.4–63.4), bad housing type (OR:2.5 [1–6.8]), partner with multiple sexual relationship (OR:4.5 [0.9–22.9]) and single women (widowed, divorced, separated, never married) (OR:6.9 [1.1–42.2]). This study provides the first national-based estimate of HPV prevalence in Tunisia. Our findings contribute to the evidence on the current burden of HPV infection, the critical role of sexual behaviour and socioeconomic status and call for increased support for the screening program in Tunisia to prevent cervical cancer. These results allow us to evaluate the cost-effectiveness of vaccine program implementation in Tunisia in future.

Clinical and analytical performance of the BD Onclarity™ HPV assay for detection of CIN2+ lesions on SurePath samples

BackgroundThe novel BD OnclarityTM HPV assay (Onclarity) on the BD Viper™ LT system (BD Diagnostics, Sparks, MD), detects E6/E7 DNA from 13 high-risk HPV genotypes and HPV66. We compared the analytical and clinical performance of the Onclarity Assay to that of Hybrid Capture 2 and LINEAR ARRAY using adjudicated histological outcomes from Danish women referred for colposcopy. Methods276 women from Copenhagen, Denmark were referred for colposcopy with abnormal cytology and/or a positive HPV test. Two samples for HPV analysis were taken in BD SurePath™ and in the BD cervical brush diluent (CBD) media. ClinicalTrial gov. identifier: NCT01671462, Ethical Approval: H-4-2012-070. ResultsHistology was normal in 84 (31%) women, 70 (26%) had CIN1, 47 (17%) CIN2, and 68 (25%) had CIN3. The Onclarity assay detected 67 out of 68 (99%) ≥CIN3 and 113/115 (98%) ≥CIN2. The specificities for <CIN2 were 21%, 17%, and 22%, for HC2, Onclarity and LA, respectively. ConclusionOverall, the Onclarity HPV assay performed well on SurePath LBC and CBD media, with clinical sensitivity and specificity matching those of HC2 and LA.

Prevalence of Human Papilloma Virus Infection in Young Primiparous Women During Postpartum Period: Study from a Tertiary Care Center in Northern India

Assessment of high-risk Human Papilloma Virus (HPV) prevalence is important for monitoring long-term decrease in cervical cancer after implementation of the prophylactic HPV vaccination. To determine the prevalence of high-risk HPV infection and cytological abnormalities in young primiparous women in the age group of 16-26years. In this cross-sectional study, 214 primiparous women aged 16-26years were recruited from a public tertiary health care center postpartum clinic between June 2013 and May 2014. Cytological analysis was performed by Pap smear test and patients underwent sampling with cervical brushes for HPV-DNA detection and typing by a PCR-based assay for HPV types 16, 18, 33 and 45. High-risk HPV was detected in 41 (19.2%) women. HPV 16 was found to be most prevalent with 17 (7.9%) samples testing positive, followed by HPV 18 in nine (4.2%), HPV 45 in six (2.8%) and HPV 31 in four (1.8%) women. Five women tested positive for more than one HPV types. There were no cases of intraepithelial lesions or cervical cancer. One patient who had Atypical Cells of Undetermined Significance (ASCUS) on cytology tested negative for all four HPV genotypes. This study provides a geographic baseline data of high-risk HPV prevalence in young Indian women before implementation of a vaccination program. The results are important for comparison with other global regions and monitoring the effect of HPV vaccination.

Cervical human papillomavirus infection and persistence: a clinic-based study in the countryside from South Brazil

Human papillomavirus (HPV) infection is common in sexually active women and viral persistence may cause intraepithelial lesions and eventually progress to cervical cancer (CC). The present study aimed to investigate epidemiological factors related to HPV infection and to evaluate viral persistence and CC precursor lesions frequencies in women from a city in the countryside of South Brazil. Three hundred women were recruited from a primary public health care clinic. The patients were interviewed and underwent sampling with cervical brushes for HPV-DNA detection/typing by a PCR-based assay and cytological analysis by Pap smear test. HPV was detected in 47 (15.7%) women. HPV infection was significantly associated with young age (<30 years) and low socio-economic status. Seventeen (5.7%) women presented cytological abnormalities, three of them with precursor CC intraepithelial lesions. A subgroup of 79 women had been previously analyzed and thirteen (16.4%) were persistently infected, two with precursor CC intraepithelial lesions and high-risk HPV types infection (both of them without cervical abnormalities in the first exam). In conclusion, HPV infection was associated with young age (<30 years) and low family income; viral persistence was low (16.4%) but related to CC precursor lesions; and HPV-DNA high risk types detection would help to screen CC in the population.

Clinical Study of Styping Detection of Human Papillomavirus (HPV) Infection with Microarray from Paraffinembedded Specimens of Cervical Cancer and Precursor Lesions.

The prevalence and type distribution of human papillomavirus (HPV) in cervical cancer and cervical intraepithelial neoplasia (CIN) in Jiangsu, China was investigated. A total of 93 cases with cervical cancer and 176 CINII-III tissue samples were obtained from women undergoing biopsy or surgery. The 1047 exfoliated cervical cell samples were collected with cervical brush in physical examination women. HPV DNA and typing were examined by polymerase chain reaction (PCR) and gene-chip. The results showed that HPV DNA was detected in 82 cases with cervical cancer (88.17%), HPV 16 being detected in 65 (69.89%) cases, HPV 18 in 12 (12.90%) cases, HPV 33 in 10 (10.75%) cases, HPV 31 in 4 (4.30%) cases, and HPV 45 in 3 (3.23%) cases. HPV DNA was detected in 154 cases with CINII-III (87.50%), HPV 16 being detected in 92 (52.27%) cases, HPV 18 in 50 (28.41%) cases, HPV 33 in 25 (14.21%) cases, HPV 58 in 25 (14.21%) cases, and HPV 31 in 20 (11.36%) cases. About 20.43% cervical cancer and 38.64% CINII-III specimens exhibited multiple infections (p < 0.01). The total positive rate, single infection and mixed infection rate of the CINII-III and SCC group all had a significant difference (p < 0.05) when compared with the normal cells group. The total positive rate, single infection rate and mixed infection rate of CINII-III group did not show significant difference (p > 0.05) when compared with SCC group. CINII-III and SCC had all intimate relation with HPV infection. The high prevalence of HPV 16, 18, 33, 31 and 58 in Jiangsu (China) deserves more attention, as it has important implications for the successful use of HPV vaccine and choice of diagnostic methods.

Prevalence of microorganisms co-infections in human papillomaviruses infected women in Northern China

Due to scarce data on the prevalence of genital microorganism infections among individuals with human papillomavirus (HPV) infections, the present study aimed to evaluate microorganism co-infections and associated risk factors in HPV-infected women in Northern China. Cervical samples of 4290 enrolled female patients were collected to detect HPV, bacterial and yeast infections in gynecologic outpatients. Serum samples collected were analyzed for the presence of serological markers for human immunodeficiency virus (HIV), hepatitis B (HBV) and hepatitis C (HCV) and Treponema pallidum infections. HPV typing was carried out by polymerase chain reaction and flow-through hybridization on cervical brush specimens. Chlamydia trachomatis was identified by strand displacement amplification, and other microorganisms were detected by conventional methods. Odds ratio of co-infection was assessed between HPV-positive and -negative patients. Overall, co-infections among HPV-infected patients showed a trend for risk compared with HPV-negative patients. In this cross-sectional study on 4290 female outpatients, statistical analyses revealed a significant association between HPV and detection of anti-HBV antibodies, bacterial vaginosis, C. trachomatis and Ureaplasma urealyticum; no correlation was found between HPV infection and anti-HIV, anti-HCV, T. pallidum, Trichomonas vaginalis and Neisseria gonorrhoeae, which were detected only in few cases among either HPV-positive or -negative patients. The study demonstrated a significant association between HPV and HBV, bacterial vaginosis, C. trachomatis and U. urealyticum. The study results suggest that it may be important to screen for the simultaneous presence of different microorganism co-infections with HPV that may have synergistic pathological effects.

Manual liquid based cytology for Pap smear preparation and HPV detection by PCR in Pakistan.

This study was conducted on female patients with different gynecological problems attending the gynecology out-patient departments of two tertiary care hospitals in Peshawar city of Khyber Pakhtunkhwa, Pakistan between August 2012 and October 2013. The 200 patients had an age range of 21-65 years. Smears were taken with cervical brushes and preserved in preservative medium and processed for manual liquid based cytology (MLBC) for Pap staining. Out of 200 collected samples, 30 samples were found inadequate on cytology. Of the remaining 170 samples, 164 (96.47%) were normal, 5 (2.94%) were of atypical squamous cells of unknown significance (ASCUS) and 1 (0.6%) was of high grade squamous intraepithelial lesion (HSIL). On PCR all the samples were positive for beta globin gene fragment including those reported inadequate on cytology. Out of the 5 ASCUS samples, 2 samples were positive for HPV, one each for HPV 16 and HPV 18, and the rest of the 3 samples were negative for HPV DNA. The 1 sample of HSIL was positive for HPV 16 on PCR. Out of 164 normal samples on cytology, only 1 sample was HPV 16 positive. So overall, 4 (2%) out of 200 samples were positive for HPV DNA, where 3 were HPV 16 (1.5%), and 1 was HPV 18 (0.5%) positive, and thus the ratio of infection with of HPV 16 to HPV 18 was 3:1 in the general population. In conclusion, PCR based HPV detection is a more sensitive method for screening of HPV infection than cytology as sample inadequacy does not affect the results. However, it can be combined with cytology methods in a HPV positive female to achieve the maximum results.

Comparative scanning electron microscopy evaluation of Canal Brushing technique, sonic activation, and master apical file for the removal of triple antibiotic paste from root canal (in vitro study).

Aims:To compare and evaluate the effectiveness of Canal Brushing technique, sonic activation, and master apical file (MAF) for the removal of triple antibiotic paste (TAP) from root canal using scanning electron microscopy (SEM).Materials and Methods:Twenty-two single rooted teeth were instrumented with ProTaper up to the size number F2 and dressed with TAP. TAP was removed with Canal Brush technique (Group I, n: 6), sonic (EndoActivator) (Group II, n: 6), and MAF (Group III, n: 6). Four teeth served as positive (n: 2) and negative (n: 2) controls. The roots were split in the buccolingual direction and prepared for SEM examination (×1000) at coronal, middle, and apical third. Three examiners evaluated the wall cleanliness.Statistical Analysis:Statistical analysis was performed by Kruskal–Wallis test and Wilcoxon rank sum test.Results:Difference in cleanliness between three groups is statistically significant in cervical region only. Pairwise comparison in cervical region Canal Brush and sonic activation showed more removal of TAP than MAF.Conclusions:Canal Brush and sonic activation system showed better result than MAF in the cervical and middle third of canal. In the apical third, none of the techniques showed a better result. None of the techniques showed complete removal of TAP from the canal.

English

AIM: Aim of our study to analyze the prevalence of premalignant lesion based on Pap smear and liquid based cytology in patients of bad cervix. OBJECTIVE: To compare sensitivity and specificity of two methods for screening of bad cervix and to know IDR (increase detection rate) of cervical cancer by conventional Pap smear and liquid based cytology. MATERIAL AND METHOD: 200 women attending Gynaecology OPD were random selection on the basis of inclusion criteria. All 200 selected women were subjected for down staging through per speculum examination for identification of bad cervix. Pap smears of all 200 selected women were taken and ensured that no local douche, antiseptic cream and no local internal examination was done on the day of test. The prepared smears were then stained according to Papanicolaou's technique. Liquid based cytology smears preparing by using cervical brush 1-1.5cm were inserted into the cervical os until the large outer bristles of the brush touch ectocervix. Data collected for sociodemographic, parity, down staging clinical examination Pap smear and LBC was organised, interpreted and analysis on appropriate statistical software. P value < 0.05 is considered significant RESULT: Analysis revealed that the maximum number of women in our study are from middle age group(31-40yrs), low socioeconomic status, married before 18 yrs, multipara, do not use any contraceptive, uneducated, urban, Muslim population.in this study more abnormal smear is seen in LBC as compared to pap smear. Sensitivity and specificity for LBC is more as compare to pap smear. CONCLUSION: In low resource setting like ours were facilities for radiology, chemo- radiotherapy and supportive care are limited or unavailable. It is important to identify which resources fill healthcare need most effectively and to consider alternative approaches, LBC is strongly advocated in the best interest of public health, by improving the quality of the sample and reducing the likelihood of false negative cytology result. Thus it will significantly improve early detection and treatment of cervical lesions and

Clinical forensic sample collection techniques following consensual intercourse in volunteers – Cervical canal brush compared to conventional swabs

The purpose of the research was to evaluate gynecological evidence collection techniques; the benefit of cervical canal brush sample compared to vaginal fornix and cervical swab samples and the time frame for detecting Y-chromosomal material QiAmp DNA Mini Kit® and Quantifiler Y Human Male DNA Quantification Kit® in adult volunteers following consensual intercourse.Eighty-four adult female volunteers following consensual intercourse were recruited for the study. By combining all sample collecting techniques, 81.0% of the volunteers were Y-DNA positive. Up to 60 h the conventional swab sampling techniques detected more Y-DNA positive samples when compared to the brush technique. However, after 60 h, the cervical canal brush sample technique showed its benefit by detecting 27.3% (6/22) of Y-DNA positive samples, which were Y-DNA negative in both conventional swab sampling techniques. By combining swab and brush techniques, 75% of the volunteers were still Y-DNA positive in 72–144 post-coital hours. The rate of measurable Y-DNA decreased approximately 3% per hour. Despite reported consensual intercourse, 6.8% (3/44) of volunteers were Y-DNA negative within 48 h. Y-DNA was not detected after 144 post-coital hours (6 days).In conclusion, the brush as a forensic evidence collection method may provide additional biological trace evidence from the cervical canal, although the best biological trace evidence collection can be obtained by combining all three sampling techniques. The time frame for gynecological forensic evidence sample collection should be considered to be at least a week if sexual violence is suspected.

PAX1 methylation as a potential biomarker for cervical cancer screening.

DNA methylation is a potential biomarker for early cancer detection. Previous studies suggested that the methylations of several genes are promising markers for the detection of cervical intraepithelial neoplasia at grade III or worse (CIN3+). The purpose of the present study was to explore the feasibility of these DNA methylation testing in cervical cancer screening. A total of 443 women were recruited from the Yuan's General Hospital. Cervical scrapings were collected for Papanicolaou (Pap) test by using cervical brushes, and the cytological data were used for analysis. The residual cells on the brush were preserved in phosphate-buffered saline solution at 4°C until DNA extraction. Then, the extracted DNA were used for molecular tests, which included human papillomavirus typing and quantification of the methylation levels for PAX1, SOX1, and NKX6-1 genes. Subjects who had abnormal Pap test results underwent colposcopy or biopsy with subsequent conization or major surgery when biopsy results revealed CIN2+. The final diagnosis for this group was confirmed by colposcopy or pathological examination. The study was approved by the institutional review board of Yuan's General Hospital, and all the molecular tests were performed by ISO17025 certified laboratories. The sensitivity of PAX1 and SOX1 was greater than 80%, and the specificity of PAX1 and NXK6-1 was greater than 80% for the detection of CIN3+ lesions. PAX1 detection alone had a sensitivity and specificity of 86% and 85%, respectively, whereas when used as a cotest with the Pap test, the sensitivity and specificity were 89% and 83%, respectively. PAX1 showed great potential as a biomarker for cervical cancer screening. When incorporating PAX1 detection into current screening protocol, the efficacy of screening could be greatly improved. Moreover, unnecessary referral for colposcopy and biopsy could be reduced up to 60%. However, prospective population-based studies are necessary for further implementation of this screening program.

Distribution of serum antibodies against human papillomavirus 16 and 18 among high-risk women to cervical cancer

To explore the distribution of serum antibodies against human papillomavirus (HPV) 16/18 among women at high-risk for cervical cancer. All women when tested positive for anyone of the cervical cancer screening programs, from Xinmi county of Henan province in 2011, were recruited as the subjects of this study. Cervical exfoliated cells were collected, using cervical brush for HPV DNA testing, and 10 ml venous blood was drawn for HPV-16, 18 serum antibodies testing, by enzyme-linked immunosorbent assay. Among the 952 women under study, 230 cases (24.2%)showed HPV DNA positive, with positivity rates of HPV16 and 18 L1 virus-like particle (VLP)antibodies as 23.2% and 6.5%, respectively. The overall positivity rate of any type of HPV16, 18 VLP antibodies was 26.8% . Geometric means of HPV16, 18 VLP antibody titers were 79.1 (Yangshengtang Unit,YU/ml) and 125.0(YU/ml). Positivity rate of HPV16 antibody was significantly associated with age, viral load of HPV DNA, and cervical lesion severity (P < 0.05). Seropositivity of HPV18 was also increasing with the increase of viral load (P < 0.01) with different cervical lesion significantly showing different titer of HPV18 antibody (P < 0.01). Based on the results of HPV DNA detection among the two years of study, women with HPV persistent infection showed significant higher positive rate of HPV16/18 antibodies than women who did not have HPV infection or emerging infection (P < 0.001). When comparing to those women without HPV infection, the ones with transient infection showed higher seropositivity rates on both HPV16 antibodies and titer of HPV16 antibody (P < 0.001). Seroprevalence rates on HPV16 and 18 among the unvaccinated high-risk women in Henan were high. Prevalence of both HPV16 and 18 antibodies were correlated with age, viral load, cervical lesion and history of infection.Women with high viral load, high grade cervical lesion or history of infection would more likely to be seropositive.

Human papillomavirus status of head and neck cancer as determined in cytologic specimens using the hybrid-capture 2 assay

A standardized assay to determine the HPV status of head and neck squamous cell carcinoma (HNSCC) specimens has not yet been established, particularly for cytologic samples. The goal of this study was to determine whether the hybrid capture-2 (HC-2) assay, already widely used for the detection of high risk HPV in cervical brushings, is applicable to cytologic specimens obtained from patients with suspected HNSCCs. Fine needle aspirates (FNA) of cervical lymph nodes were pre-operatively obtained from patients with suspected HNSCCs and evaluated for the presence of HPV using the HC-2 assay. HPV analysis was performed on the corresponding resected tissue specimens using p16 immunohistochemistry (IHC) and HR-HPV in situ hybridization (ISH). A cost analysis was performed using the Center for Medicare & Medicaid Services. HPV status of the cervical lymph node metastases was correctly classified using the HC-2 assay in 84% (21/25) of cases. Accuracy was improved to 100% when cytologic evaluation confirmed the presence of cancer cells in the test samples. The estimated cost savings to CMS using the HC-2 assay ranged from $113.74 to $364.63 per patient. HC-2 is a reliable method for determining the HPV status of HNSCCs. Its application to HNSCCs may reduce costs by helping to localize the primary site during the diagnostic work-up as well as decrease the interval time of determining the HPV status which would be relevant for providing prognostic information to the patient as well as determining eligibility for clinical trials targeting this unique patient population.

Chlamydia trachomatis infection and human papillomavirus in women with cervical neoplasia in Pernambuco-Brazil

Chlamydia trachomatis (CT) is the most common bacterial cause of sexually transmitted disease. High-risk human papillomavirus (HR-HPV) is considered the main etiological agent for cervical neoplasia. Evidences showed that the presence of co-infection of CT and HR-HPV plays a central role in the etiology of cervical intraepithelial neoplasia (CIN) and cervical cancer. The goals of this study were: evaluate the human papillomavirus (HPV) and CT prevalence among Brazilian women with abnormal cytology and provide the effect of this association on the severity of cervical neoplasia. The population of this study was composed by 142 women with incident histological incidence of CIN grades I, II, III or cervical cancer from Recife, Northeast of Brazil. The polymerase chain reaction method on a cervical brush specimen was used to detect both agents and the automatic sequencing method was used for HPV genotyping assay. The prevalence of HPV and CT was 100 and 24.65%, respectively. Thirteen types of HPV were detected; HPV 16, 18, 31 and 33 were the most common. The most prevalent HPV types were HPV 16 and 18. A significant association between CT positive and HPV 16 infection was found (p<0.0106; OR=5.31; 95% IC1.59-17.67). In the study population, there was diversity of HPV infections, with high-risk types being the most common. Also, the data collected suggest that CT infection may play an important role in the natural history of HPV infection.

Urine-based human papillomavirus DNA testing as a screening tool for cervical cancer in high-risk women

ObjectiveTo test the hypothesis that self-collected urine could be used to detect high-risk human papillomavirus (HPV) DNA with sensitivity and specificity comparable to those of standard cervical testing. MethodsWomen attending a gynecology clinic for evaluation of abnormal cytology were recruited. Fifty-two participants (21–60years of age) collected urine samples, and clinicians collected cervical brush samples. When appropriate, cervical biopsies were obtained during colposcopy. HPV detection and typing were performed on DNA extracts from each sample, using commercial reagents for L1 consensus polymerase chain reaction (PCR) and type-specific hybridization. HPV 16 viral load was determined by quantitative PCR in HPV 16-positive samples. A diagnostic test analysis was conducted for urine samples. ResultsFifty paired samples were analyzed, with 76% agreement between samples. The 12 discrepant pairs were all urine negative/cervix positive. The most common HPV types detected were 16, 51, 53, and 62. The urine test correctly identified 100% of the uninfected and 65% of the infected patients. ConclusionThe results indicate that HPV DNA detection using urine is less sensitive than cervical sampling in a population with abnormal cytology. Further exploration is warranted to determine clinical utility when other options are unavailable.

The prevalence and risk factors of human papillomavirus in female sex workers.

Human papillomavirus virus (HPV) is the major causative factor for cervical cancer, and sex workers are at high risk for HPV infection. In this study, we aimed to estimate the prevalence and risk factors of HPV infection among female sex workers (FSWs). The study included 239 brothel-based FSWs who work in Izmir, Turkey. A self-administered questionnaire for risk factors was completed, and cervical brush samples were taken for HPV detection and typing. HPV detection and typing were performed by multiplex polymerase chain reaction (PCR) and reverse hybridization methods. The risk factors related to HPV infection were determined by multivariate analysis. The prevalence of HPV among FSWs was 20.1%. HPV18 was the most common type (40%), followed by HPV16 (17%) and HPV50 (15%). Logistic regression analysis revealed that being less than 30 years of age, having a high frequency of sexual contacts, smoking, and lack of condom use were significantly associated with HPV infection. FSWs have a high prevalence of HPV infection and are at increased risk for cervical cancer. As they are a priority group for active follow-up, national strategies for reducing HPV among FSWs and regular cervical cancer screening programs should be implemented for this population.

Maternal transmission of human papillomavirus in retinoblastoma: A possible route of transfer

Context:After establishing the presence of human papillomavirus (HPV) in retinoblastoma (RB), the probable role of the mother was investigated.Materials and Methods:A total of 21 sporadic RB cases and 15/21 corresponding mothers′ cervical brushings were collected. HPV testing was carried out using multiplex PCR (PGMY09/11 primers) followed by genotyping using line blot assay.Results:We found both high- (83%) and intermediate-risk (17%) HPV types in 12/21 (57%) RB samples and only high-risk (100%) types in 6/15 (40%) cervical brushing samples. The single genotype of HPV 16 was found in six cases and HPVs 82, 68 and 35 in one case each. Both HPVs 16 and 59 were found in two cases and HPV 16 and 73 in one case. Three samples of RB harboring HPV 16, HPVs 16 and 59, and HPVs 16 and 73 had HPV genotype 16 in the respective mothers′ cervical brushing samples.Conclusions:Maternal transfer of HPV in RB could be a possible route of transmission. However, a larger cohort and sampling of the mothers′ cervical brushings at various stages, i.e. before, during, and after pregnancy will give us insight to propound an alternate mechanism for the development of sporadic RB.