Aaron Ermel

Human papillomavirus detection and typing in thin prep cervical cytologic specimens comparing the Digene Hybrid Capture II Assay, the Roche Linear Array HPV Genotyping Assay, and the Kurabo GeneSquare Microarray Assay

Three methods for the detection of HPV DNA were compared in cervical cytologic specimens: the Digene Hybrid Capture II Assay (HC), the Roche Linear Array HPV Genotyping Assay (LA) and the Kurabo GeneSquare Microarray (GS). The main goals of the study were to correlate cytology with HPV detection and to determine agreement between assay pairs for HPV detection. Thin-prep Pap smears were performed and supernates were tested by HC, LA, and GS. For specimens reacting with the HPV 52/33/35/58 probe in the LA assay, type-specific PCR was performed for HPV types 52, 33, 35, or 58. Binomial proportions and kappa coefficients were calculated for agreement between assays. Cytology results and supernatant were available for 202 subjects. HPV detection increased with worsening cytologic abnormality in all three assays. For all cytologic groups, LA and GS detected more HPV (all and oncogenic) than HC. However, for detection of oncogenic HPV types represented in all three assays, differences between assays were less pronounced. The highest agreement was between LA and GS. In four of 12 specimens reacting with the HPV 52/33/35/58 probe in the LA assay but deemed HPV 52-LA-negative using an algorithm provided by the manufacturer, the presence of HPV 52 was confirmed using type-specific HPV 52 PCR. All four of these specimens were also GS-positive for HPV 52.

Association of Chlamydia trachomatis Infection With Redetection of Human Papillomavirus After Apparent Clearance

BACKGROUND Persistent infection with oncogenic human papillomavirus (HPV) is associated with an increased risk of cervical malignancy. Redetection of type-specific HPV after a period of nondetection may be caused by reactivation of a low-level persistent infection. Little is known about factors associated with type-specific HPV redetection. METHODS For a longitudinal cohort of adolescent women with frequent behavioral and sexually transmitted infection (STI) information (every 3 months), Cox proportional hazard models were used to assess the influence of sexual behaviors and STIs on the redetection of oncogenic or high-risk HPV infections. RESULTS A total of 210 type-specific high-risk HPV detection episode periods were identified in this longitudinal cohort; 71 (33.8%) were characterized by a period of nondetection followed by redetection. Chlamydia trachomatis (hazard ratio [HR], 3.14; 95% confidence interval [CI], 1.44-6.86) was associated with redetection; redetection was >2 times more likely with each additional self-reported sex partner in the past 3 months (HR, 2.26; 95% CI, 1.35-3.78). CONCLUSIONS This study demonstrates the role of C. trachomatis and number of recent sexual partners in type-specific HPV redetection. Given that persistent oncogenic HPV infections are associated with cancer-related outcomes, understanding the potential role of such factors in the pathogenesis of HPV-related outcomes is important.

Low-level persistence of human papillomavirus 16 DNA in a cohort of closely followed adolescent women†

Most human papillomavirus (HPV) infections in young women become undetectable by standard assays after a few months. It is possible that many HPV infections do not actually clear, but persist at very low levels for years, becoming detected again later in life. The purpose of this study is to describe HPV 16 clearance, reappearance, and low‐level persistence in a cohort of adolescent women. Adolescent women (N = 66), not vaccinated against HPV, were recruited from 1998 to 2008 into a longitudinal study. Self‐collected vaginal samples were obtained quarterly and tested for HPV by Linear Array HPV Genotyping Test (LA‐HPV). To explore low‐level persistence, a type‐specific nested PCR for HPV 16 (TSN‐PCR‐16) was developed. Women with HPV 16 detected by LA‐HPV had their negative swabs retested with TSN‐PCR‐16. Forty‐two participants with HPV 16, followed for a mean of 6.3 years, were analyzed. Using LA‐HPV, the median duration of HPV 16 detection was 428 days (SD 852.5 days). TSN‐PCR‐16 detected HPV 16 during periods of LA‐HPV non‐detection in samples from many women. Using a combination of LA‐HPV and TSN‐PCR‐16 results, the median duration of HPV 16 detection was 1,022.5 days (SD 943.7 days). The durations of detection differed significantly between the two methods (P = 0.0042) with a mean difference of 434.5 days. In adolescent females, duration of HPV 16 detection was significantly longer when TSN‐PCR‐16 was combined with LA‐HPV. Some apparently cleared HPV 16 could be shown to persist at low levels using nested PCR. J. Med. Virol. 83:1362–1369, 2011.

Episodic detection of human papillomavirus within a longitudinal cohort of young women

Redetection of a type‐specific human papillomavirus (HPV) infection may represent reinfection. However, a growing body of literature suggests that reactivation of HPV is common and that episodic detection of a HPV infection may represent reactivation of a persistent virus. A cohort of prospectively followed adolescent women (N = 150), ages 14–17, was observed on average 6.4 years. The authors describe the redetection of 37 HPV types and associated factors of redetection of high‐risk (HR) and low‐risk (LR) types using Cox proportional hazard models. Of 1,248 HPV type‐specific infections, 286 (22.9%) were associated with redetection after apparent clearance. Chlamydia infections (HR = 1.99 [95%CI, 1.15–3.49]) and non‐condom use (HR = 1.1 [95%CI, 1.04–1.99]) were associated with increased redetection of HR‐HPV infections. Oral contraceptive pills (HR = 2.73 [95%CI, 1.52–4.90]) and number of sexual partners (HR = 1.44 [95%CI, 1.04–1.99]) were associated with increased redetection of LR‐HPV infections. Episodic detection of HPV is common for HR‐ and LR‐HPV types. This finding and identified factors or redetection have clinical implications and enhances the understanding of HPV natural history. J. Med. Virol. 87:2122–2129, 2015.

Invasive cervical cancers from women living in the United States or Botswana: differences in human papillomavirus type distribution.

BackgroundCervical cancer is the primary cause of cancer-related deaths in women living in Botswana.MethodsParaffin-embedded blocks of formalin-fixed invasive cervical cancer specimens were identified from women living in the U.S. (n = 50) or Botswana (n = 171) from which DNA was extracted. Thin-section PCR was performed on each sample for HPV types and HIV. Comparisons were made between HPV types and groups of types identified in cancers.ResultsHPV DNA was identified in 92.0% of specimens from the U.S. containing amplifiable human DNA, and 79.5% of specimens from Botswana. HPV 16 was detected in 40 of 46 HPV-positive specimens (87.0%) from the U.S. vs. 58 of 136 (42.7%) from Botswana (p < 0.001). In contrast, non-HPV 16/18 types, all A9 species (HPV16, 31, 33, 35, 52, and 58), non-HPV 16 A9 (HPV 31, 33, 35, 52, and 58), HPV 18, all A7 types (18, 39, 45, 59, and 68) types were detected significantly more often in specimens from Botswana. The prevalence of non-HPV 18 A7 types did not differ significantly between the two groups. For specimens from Botswana, 31.6% contained PCR-amplifiable HIV sequences, compared to 3.9% in U.S. specimens. Stratifying the samples from Botswana by HIV status, HPV 31 was detected significantly more often in HIV-positive specimens. Other HPV types and groups of types were not significantly different between HIV-positive and HIV-negative specimens from Botswana.ConclusionThis study demonstrates that there may be important HPV type differences in invasive cervical cancers occurring in women living in the United States or Botswana. Factors in addition to HIV may be driving these differences.

DNA detection and seroprevalence of human papillomavirus in a cohort of adolescent women.

Objectives Human papillomavirus (HPV) infections are common in adolescent women, while the rare cancerous sequelae of HPV infections do not generally occur until the 4th or 5th decades of life. This prospective study of a cohort of adolescent women was performed to further our knowledge of the natural history of incident and prevalent HPV infections. Methods Self-vaginal swabs collected from high-risk, unvaccinated adolescent women in a longitudinal study were analysed for HPV DNA. Sera were collected at enrolment and later tested for HPV antibodies. Statistical analysis was performed to determine the HPV genotype distribution and duration of detection, and to determine rates of seropositivity and seroconversion for HPV types represented in the assays. Results 146 subjects (mean enrolment age=15.4 years; mean duration of follow-up=5.8 years) had samples adequate for analysis of HPV detection, and 95 of these subjects had paired sera available. The cumulative prevalence for high-risk and low-risk HPV types was 95.9% and 91.1%, respectively. HPV types 6, 11, 16 and 18 (HPV types represented in the quadrivalent vaccine) were found at some point in 40.4%, 6.2%, 48% and 24% of participants, respectively. Serological data confirmed exposure to these vaccine-covered types, as well as to other high-risk HPV types. Conclusions In this cohort of adolescent women, high- and low-risk HPV types were frequently detected, and serological data confirmed exposure in most subjects. The high-prevalence HPV types represented in the quadrivalent HPV vaccine further support vaccination of women at an age well before sexual debut.

Redetection of human papillomavirus type 16 infections of the cervix in mid-adult life

Purpose To assess whether HPV 16 originally detected in adolescent women can be redetected in adulthood. Methods A convenience sample of 27 adult women with known HPV 16 detection during adolescence was assessed for HPV 16 redetection. A comparison of the long control region (LCR) DNA sequences was performed on some of the original and redetected HPV 16 isolates. Results Median age at reenrollment was 27.5 years (interquartile range of 26.7–29.6). Reenrollment occurred six years on average after the original HPV 16 detection. Eleven of 27 women had HPV 16 redetected. Some of these HPV 16 infections had apparently cleared during adolescence. LCR sequencing was successful in paired isolates from 6 women; in 5 of 6 cases the redetected HPV 16 isolates were identical to those detected during adolescence, Conclusions HPV 16 may be episodically detected in young women, even over long time periods. HPV 16 redetection with identical LCR sequences suggests low-level persistent infection rather than true clearance, although newly acquired infection with an identical HPV 16 isolate cannot be excluded. However, this study suggests that a new HPV 16-positive test in a clinical setting may not indicate a new infection.

P3.120 Prevalence of ureaplasma urealyticum in urine of men attending a sexually transmitted disease clinic

Introduction Ureaplasma urealyticum (UU) is probably one of the causes of non-gonococcal, non-chlamydial urethritis in men and post-partum endometritis in women. The epidemiology of UU is currently unclear because culture isolation is difficult and molecular identification is limited to specialised laboratories. Testing for UU would be useful for surveillance, disease management, and epidemiology. This study assessed the prevalence of UU in men attending the local STD clinic by real-time PCR. Methods A convenience sample of de-identified residual urine specimens from men attending an STD clinic was collected. Urine was placed into commercially available transport tubes and tested by PCR for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Trichomonas vaginalis (TV). The remaining residual processed specimen was tested for UU DNA using a previously published real-time PCR assay. Descriptive statistics were used to examine UU prevalence with age, and co-infection with CT, NG, and TV. Results A total of 99 residual male urine specimens were available for testing. UU DNA was detected in 16/99 (16.2%) of the specimens and was comparable to CT (14/97, 14.4%), NG (11/97, 11.3%), and TV (4/25, 16.0%). Of the 16 UU positive specimens, co-infection with CT was observed in one (6.25%), NG in one (6.25%), and the remaining 14 (87.5%) had no other infection identified. The mean age of those individuals positive for UU DNA was 32.4 (range 18–63) while the mean age of those infected with CT, NG, and TV was 29.8, 28.3, and 36.8 years old, respectively. Conclusions The prevalence of UU in men attending an STD clinic is similar to that observed for CT, NG, and TV. This study was useful in order to gain a better understanding of UU in this population including the age of those positive for UU, co-infection status with other commonly identified pathogens, and as a means to confirm the feasibility of real-time PCR testing using residual processed specimens. More studies are needed to elucidate the significance of UU in this population.

Behavioral Correlates of HPV-Associated Oropharyngeal Squamous Cell Carcinomas

There is a well-recognized subset of head and neck squamous cell carcinomas (HNSCC) that are associated with HPV infection. These cancers differ from traditional HNSCC in that they occur in younger individuals and have decreased association with tobacco and alcohol use. Many studies have looked at the behavioral correlates associated with this subset of cancers. The most commonly reported sexual behaviors associated with this subset of HNSCC include increased number of lifetime vaginal or oral sex partners, younger age at sexual debut, oral-anal sex partners, history of other sexually transmitted infections and same-sex sexual partners. Nonsexual behaviors including tobacco use, alcohol use, marijuana use, nutrition and dental health have less consistent data supporting association with this subset of HNSCC. Understanding of the behavioral factors contributing to oral HPV DNA detection and development of HPV-associated HNSCC has increased significantly over the years, but further studies are needed to delineate the main contributors to develop effective interventions.