Cervicovaginal Swabs Research Articles

Sustained maternal antibody and cellular immune responses in pregnant women infected with Zika virus and mother to infant transfer of Zika-specific antibodies.

Evaluation of Zika virus (ZIKV)-specific humoral and cellular immune response in pregnant women exposed to ZIKV. In this observational, prospective cohort study, we recruited pregnant women presenting for prenatal ultrasound for ZIKV exposure at a single academic teaching hospital in Boston, MA from November 2016 to December 2018. We collected blood, urine, and cervicovaginal swabs antepartum, intrapartum, and postpartum; and cord blood and placenta at delivery. We used experimental assays to calculate quantitative viral loads, ZIKV-specific immunoglobulin titers, and ZIKV-specific T-cell responses. We enrolled 22 participants, three of which had serologic-confirmed ZIKV infection. No participants demonstrated sustained ZIKV shedding. ZIKV-specific IgG/IgM antibody was sustained throughout pregnancy and postpartum. ZIKV envelope and capsid-specific T-cell responses were also observed, albeit inconsistent. No newborns in this cohort had congenital Zika syndrome. Infant cord blood of infected mothers exhibited ZIKV-specific IgG, but not IgM antibodies. We detected a robust, prolonged maternal humoral immune response to ZIKV during pregnancy and postpartum. We also demonstrated evidence for efficient transplacental antibody transfer from mother to infant at birth, supporting the importance of neonatal passive immunity to ZIKV. Maternal T-cell responses were less consistent among pregnant women infected with ZIKV.

An economical Nanopore sequencing assay for human papillomavirus (HPV) genotyping

BackgroundHuman papillomavirus (HPV) testing has been employed by several European countries to augment cytology-based cervical screening programs. A number of research groups have demonstrated potential utility of next-generation sequencing (NGS) for HPV genotyping, with comparable performance and broader detection spectrum than current gold standards. Nevertheless, most of these NGS platforms may not be the best choice for medium sample throughput and laboratories with less resources and space. In light of this, we developed a Nanopore sequencing assay for HPV genotyping and compared its performance with cobas HPV Test and Roche Linear Array HPV Genotyping Test (LA).MethodsTwo hundred and one cervicovaginal swabs were routinely tested for Papanicolaou smear, cobas HPV Test and LA. Residual DNA was used for Nanopore protocol after routine testing. Briefly, HPV L1 region was amplified using PGMY and MGP primers, and PCR-positive specimens were sequenced on MinION flow cells (R9.4.1). Data generated in first 2 h were aligned with reference sequences from Papillomavirus Episteme database for genotyping.ResultsNanopore detected 96 HPV-positive (47.76%) and 95 HPV-negative (47.26%) specimens, with 10 lacking β-globin band and not further analyzed (4.98%). Substantial agreement was achieved with cobas HPV Test and LA (κ: 0.83–0.93). In particular, Nanopore appeared to be more sensitive than cobas HPV Test for HPV 52 (n = 7). For LA, Nanopore revealed higher concordance for high-risk (κ: 0.93) than non-high risk types (κ: 0.83), and with similar high-risk positivity in each cytology grading. Nanopore also provided better resolution for HPV 52 in 3 specimens co-infected with HPV 33 or 58, and for HPV 87 which was identified as HPV 84 by LA. Interestingly, Nanopore identified 5 additional HPV types, with an unexpected high incidence of HPV 90 (n = 12) which was reported in North America and Belgium but not in Hong Kong.ConclusionsWe developed a Nanopore workflow for HPV genotyping which was economical (about USD 50.77 per patient specimen for 24-plex runs), and with comparable or better performance than 2 reference methods in the market. Future prospective study with larger sample size is warranted to further evaluate test performance and streamline the protocol.

Seroprevalence of HPV Infections in the Cervix and Penis in Unvaccinated Individuals: A Study of 3,143 Participants [31F

INTRODUCTION: To determine the seroprevalence of cervical and penile HPV infections in unvaccinated, sexually-active adults in the United States. METHODS: Data were obtained from the National Health and Nutrition Examination Survey (NHANES) from 2007 to 2016. HPV serotypes were determined by cervicovaginal swabs in females and penile specimens in males. Chi-square was used for statistical analysis. RESULTS: Of 3,143 unvaccinated, sexually-active adults (median age: 28 years), 2,608 (83%) were female and 535 (17%) were male. Median lifetime sexual partners was 5 for women and 6 for men. 59.1% of females and 45.8% of males were infected with HPV. For unvaccinated women, of the top five HPV serotypes, only HPV 16 and 51 were considered high-risk and comprised 6.81% and 5.45%, respectively. The additional serotypes included HPV 53 (6.9%), 89 (6.0%), and 84 (5.7%). Of the top five HPV serotypes in unvaccinated men, HPV 51 and 66 were considered high-risk and comprised 7.2% and 5.8% of all infections; the remaining serotypes included HPV 62 (7.3%), 53 (5.5%), and 89 (5.4%). Infection with HPV serotypes covered by the 9-valent vaccine represented 21.8% of infections in females and 22.1% of infections in males in this study cohort (P=.96). CONCLUSION: In unvaccinated individuals, both men and women are at risk for infection for cancer-associated HPV types with varying seroprevalence. The efficacy of the 9-valent vaccination may have varying efficacy depending on gender and site of infection.

183: High-risk cervicovaginal microbial communities and preterm birth: a multinomial model

Community state IV (CST IV) is a high-risk cervicovaginal (CV) microbial community associated with preterm birth (PTB). Rigor of PTB phenotyping has varied among studies investigating the association of CST IV and PTB. We hypothesize that CST IV would be more strongly associated with spontaneous PTB (sPTB) than medically indicated PTB (mPTB). A prospective pregnancy cohort, Motherhood and Microbiome was performed. CV microbiota was analyzed by 16S rRNA sequencing and microbial communities were classified into community state types (CST) from CV swabs at 16-20 weeks. CV β-defensin (βD) levels were assessed by ELISA. Multinomial logistic regression was utilized to assess the association between CST IV and a 3-level birth outcome variable including sPTB, mPTB, and term birth as the reference group. The effect of immune marker βD on PTB and on the association between CST IV and birth outcome was assessed. The final cohort of 581 women included 103 sPTB, 57 mPTB, and 421 term births. In a logistic regression model adjusted for African American race and BMI, CST IV was significantly associated with all PTB (aOR 1.75; CI 1.18-2.61). In multinomial logistic regression models, CST IV was associated with sPTB (aOR 1.80; CI 1.13-2.88) and non-significantly with mPTB (aOR 1.67; CI 0.92-3.03). In adjusted multinomial models, there was no evidence of effect modification by low βD on the relationship between CST IV and sPTB (p=0.835) or CST IV and mPTB (p=0.971). The odds of sPTB for women with both CST IV and low βD were greater (aOR 3.05; CI 1.56-5.95) compared to women with neither factor (CST IV only aOR 1.80; CI 1.13-2.88 and low βD only aOR1.60; CI 1.05-2.45). CST IV increases the risk of all PTB, specifically sPTB. A high-risk CV microbial community and a poor immune response (low βD) are independent risk factors for sPTB. Further research is needed to determine if an unfavorable CV microbial-immune profile contributes to other adverse birth outcomes. Importantly, targeting the CV microbial-immune profile appears to be an important next step to reduce PTB. R01NR01478 (ME)View Large Image Figure ViewerDownload Hi-res image Download (PPT)

378: Cervicovaginal MMP-9 is associated with high-risk microbial communities and spontaneous preterm birth

Cervicovaginal (CV) microbial communities and host responses have been implicated in cervical remodeling leading to spontaneous preterm birth (sPTB). We hypothesize that disruption of the CV epithelial barrier leads to an increase in matrix metalloproteinase 9 (MMP-9), contributing to degradation of the cervical extracellular matrix, cervical shortening, and sPTB. A nested case-control was performed from a prospective pregnancy cohort, Motherhood and Microbiome which included African American women who had sPTB (n=40) or term birth (n=40). CV microbiota was characterized by 16S rRNA gene sequencing, and we classified microbial communities into community state types (CST) from CV swabs at 20-24 weeks. CST IV is a high-risk community characterized by anaerobes and a lack of Lactobacillus. Cervical length was assessed at the anatomy ultrasound with short cervix < 25mm. MMP-9 was quantified by ELISA. Standard analytical approaches and multivariable logistic regression were used to determine differences in MMP-9 and associations with outcomes. Demographic characteristics are in Table 1. MMP-9 was 4.7-fold higher in sPTB v. term birth (p=0.0013). MMP-9 was 7.3-fold higher in women with CST IV v. non-CST IV (p< 0.0001) and 4.71-fold higher in women with short cervix v. normal cervical length (p=0.01). Women with both CST IV and short cervix had a 14.5-fold increase in MMP-9 v. women with non-CST IV and normal cervical length (p< 0.0001). High MMP-9 was associated with 2.8-fold higher odds of CST IV (CI 1.05-7.63), 3.1-fold higher odds of short cervix (CI 1.05-9.17), and 3.8-fold higher odds of sPTB (CI 1.41-10.09). These findings begin to ascribe a biological link between high-risk CV microbial communities, short cervix, and sPTB through increased expression of MMP-9, which may serve to degrade the cervical matrix. Additional studies are warranted to determine the efficacy of targeting this pathway to prevent sPTB. Incorporating CV MMP-9 levels into risk stratification for sPTB may have clinical utility. (Danielle Peress MD Memorial Fund; KG & R01NR01478; ME)View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Loop-Mediated Isothermal Amplification (LAMP) for Detection of Ureaplasma diversum from Cervico-vaginal Swab of Buffaloes.

The main plan of the current study was to develop a rapid, robust, and field-applicable loop-mediated isothermal amplification (LAMP) assay for the detection of Ureaplasma diversum. A strain-specific 16S rRNA gene of Ureaplasma diversum was used for detection which was cloned, sequenced, and characterized earlier. LAMP results were visualized within 90 min with the naked eye. Cervico-vaginal swabs of 50 buffaloes were randomly collected from Livestock Research Center of NDRI as per the Institute Animal ethics guidelines. Out of 50 cervico-vaginal swab samples collected randomly, 34 were found positive with LAMP while 16 samples were negative. Conventional PCR results showed the same result. Therefore, the accuracy of the developed LAMP was about 100%. The developed LAMP assay can also be used to screen the animals for Ureaplasma diversum infection in cervico-vaginal swab. However, further study is needed to assess sensitivity and accuracy towards their detection and their relationship in disease diagnosis.

Group B Streptococcus serotypes Ia and V induce differential vaginal immune responses that may contribute to long term colonization of the female reproductive tract.

Group B Streptococcus (GBS) is a common colonizer of the female genital tract at the time of pregnancy and has been associated with severe neonatal infections. Despite trials for GBS vaccines already being underway, the factors influencing vaginal GBS colonization and clearance are currently poorly understood. Within this study, we investigated the host immune responses to GBS infections in mice that affect GBS vaginal colonization and clearance. Cervicovaginal swabs were used to measure vaginal GBS persistence, and vaginal cytokine responses were measured using the BioPlex® system. Lymphocytes isolated from spleens were stimulated with UV-killed GBS to examine systemic cellular responses. Additional in vitro cellular experiments using human vaginal epithelial cells were also performed, examining the effect pregnancy level hormones had on GBS adhesion, invasion, and cytokine responses. We observed significant differences in the ability of GBS serotype V infections to persist, compared with GBS serotype Ia vaginal infections. Vaginal cytokine response examination identified temporal changes in cytokine production (IL10, IFNγ, IL6, IL1β, and TNFα) in relation to GBS serotype and clearance or colonization. Lymphocyte proliferation assays also revealed robust cellular immune responses to GBS vaginal infections irrespective of clearance or colonization. In vitro human cellular analyses also identified that vaginal epithelial cell line cytokine production was suppressed in the presence of hormones despite no alteration in adhesion/invasion. Here, we establish previously unknown, serotype specific, temporal immune responses which may be associated with vaginal GBS colonization or clearance in the female genital tract.

HPVPro Study: Comparison of HPV Detection in Clinician-collected Cervical Swabs and Self-sampled Cervicovaginal Swabs

HPVPro Study: Comparison of HPV Detection in Clinician-collected Cervical Swabs and Self-sampled Cervicovaginal Swabs

Human miscarriage and infection in Tunisia: Role of Mycoplasma hominis and high Waddlia seroprevalence

Miscarriage is one of the most common adverse pregnancy outcomes. The aim of this study was to investigate the relationship between miscarriage in humans and infections caused by zoonotic bacteria and genital pathogens. Cervicovaginal swabs and placenta samples from 132 women with miscarriage (patient group: PG), and cervicovaginal swabs from 54 women with normal pregnancy (control group:CG), were subjected to bacteriological culture and real time PCRs detecting Coxiella burnetii, Brucella spp, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum, Chlamydia trachomatis, Waddlia chondrophila and Parachlamydia acanthamoebeae DNA. Serology of C. burnetii, C. trachomatis and W. chondrophila was also performed. Placenta samples were positive for E. coli, S. agalactiae, U. urealyticum, M. hominis and C. trachomatis in 4.7%, 3.1%, 3.1%, 0.7% and 0.7% of cases, respectively. For cervicovaginal swabs, M. hominis was more frequently detected among PG than CG with a significant statistical difference (p = 0.02). C. trachomatis was detected in 3.3% and 5.5% among PG and CG, respectively. U. urealyticum DNA was detected with high percentages in the two groups. Samples from both groups showed negatives results for C. burnetii, Waddlia, and Brucella qPCRs. A high rate of W. chondrophila seroprevalence (42%) was noted with significant difference among women with early miscarriage. C. trachomatis, S. agalactiae and M. hominis may play a role in miscarriage. However, the full characterization of the vaginal flora using other technologies such as NGS-based metagenomics is needed to clarify their role in miscarriage. Finally, further investigations should be performed to explain high W. chondrophila seroprevalence.

The Effect of Hormonal Contraception on Cervicovaginal Mucosal End Points Associated with HIV Acquisition.

Reproductive age women may choose to concurrently use topical antiretrovirals and hormonal contraceptives (HCs) to simultaneously prevent HIV-1 infection and unintended/mistimed pregnancy. There are conflicting data on the effect of HCs on mucosal susceptibility to HIV-1. The objective of this study was to evaluate cervicovaginal (CV) mucosal data from healthy women before and after initiation of either oral contraceptive pills (OCPs) or depot medroxyprogesterone acetate (DMPA) injection. CONRAD A10-114 was a prospective, open-label, parallel cohort study. We enrolled 74 women and 62 completed the visits (32 and 30 who selected OCPs and DMPA, respectively). Participants provided CV lavage, vaginal biopsies, and CV swabs at baseline in the luteal phase and then ∼6 weeks after initiating HCs. After contraceptive initiation, there were significant increases in vaginal immune cell density among both DMPA and OCP users. Changes for OCP users were concentrated in the subepithelial lamina propria, whereas for DMPA users, they were distributed throughout the vaginal tissue, including the epithelium (CD45+, CD3+, CD4+, and CD1a+). Contraceptive use altered concentrations of soluble CV inflammatory and immune mediators, with significant reductions in some proinflammatory cytokines and secretory leukoprotease inhibitor. Compared with baseline, p24 antigen production after ex vivo HIV-1 infection of vaginal biopsies doubled after DMPA use, but all p-values were >.05. HIV-1 replication was significantly higher in DMPA-exposed tissues compared with those from the OCP group at the end of the tissue culture (p = .01). Although not statistically significant, median in vitro inhibition of HIV-1 by CV fluid (innate antiviral activity), was reduced by ∼50% with HCs (p > .21). Exposure to exogenous contraceptive hormones significantly increased vaginal immune cells and reduced CV proinflammatory cytokines and antimicrobial peptides. DMPA users showed higher susceptibility to HIV-1 ex vivo infection.

Detecting Ureaplasma urealyticum among HIV-infected women with or without human papillomavirus using real-time PCR with the ANYPLEX™ II STI-7 assay system.

ObjectivesHuman immunodeficiency virus (HIV) infections increase the chances of women contracting human papillomavirus (HPV), the prime causative agent of cervical cancer. Additionally, Ureaplasma urealyticum is the most frequent pathogen prevailing in sexually transmitted diseases in HIV-infected women. The aim of this study was to determine the presence of U. urealyticum among HIV-infected women with or without HPV in Ghana.MethodsDNA samples were extracted from cervico-vaginal swabs obtained from 96 HIV-infected women attending the Kumasi South Hospital. U. urealyticum in the DNA samples was detected by real-time PCR with the Anyplex™ II STI-7 detection assay. Microsoft Excel was used to analyze the data, and the Chi-square test was used to determine associations and dependence among the variables.ResultsAmong the study population, 93.75% (90/96) were positive for at least one pathogen.In total, 36.5% (35/96) of the women were infected by U. urealyticum, with 30.21% (29/96) being co-infected with HPV. There was no significant association (95% CI, p > 0.05) between U. urealyticum and HPV status among the HIV-infected women.ConclusionU. urealyticum and HPV are highly pathogenic, and their prevalence in this study reiterates the need for their routine screening in HIV-infected patients.

Multisite HPV infections in the United States (NHANES 2003–2014): An overview and synthesis

HPV is the most common sexually transmitted infection in the U.S., infecting both anogenital and oral sites. Nationally representative data are collected through the National Health and Nutrition Examination Survey (NHANES). However, changing designations of HPV genotypes as high or low risk and varying underlying populations as new results are reported have made direct comparison of results difficult. We reanalyzed HPV data from NHANES derived from self-collected cervicovaginal swabs (women ages 18–59, 2003–14), penile swabs (men ages 18–59, 2013–14), and oral rinses (men and women ages 18–69, 2009–14), using consistent populations and definitions across NHANES cycles. These data strengthen our understanding of age trends in HPV prevalence: cervicovaginal prevalence decreases with age, penile prevalence increases with age, and oral prevalence is bimodal but with an earlier first peak in women. There is strong evidence for reduced prevalence of vaccine genotypes (6, 11, 16, 18) in vaccinated men and women (ages 18–24) at both genital (RR 0.2 (0.1–0.3) in women and 0.7 (0.1–5.4) in men) and oral sites (RR 0.1 (0.0–1.3) in women; no infections detected in vaccinated men). A more complete picture of the burden of HPV in the U.S. is emerging, including evidence for reduced HPV genital and oral prevalence in vaccinated individuals.

Defining characteristics of genital health in South African adolescent girls and young women at high risk for HIV infection.

The genital tract of African women has been shown to differ from what is currently accepted as ‘normal’, defined by a pH≤4.5 and lactobacilli-dominated microbiota. Adolescent girls and young women (AGYW) from sub-Saharan Africa are at high risk for HIV, and we hypothesized that specific biological factors are likely to be influential. This study aimed to compare characteristics of vaginal health in HIV-negative AGYW (16-22-years-old), from two South African communities, to international norms. We measured plasma hormones, vaginal pH, presence of BV (Nugent scoring), sexually transmitted infections (multiplex PCR for Chlamydia trachomatis, Neisseria gonorrhoea, Trichomonas vaginalis, Mycoplasma genitalium) and candidiasis (Gram stain) in AGYW (n = 298) from Cape Town and Soweto. Cervicovaginal microbiota was determined by 16S pyrosequencing; 44 genital cytokines were measured by Luminex; and cervical T-cell activation/proliferation (CCR5, HLA-DR, CD38, Ki67) was measured by multiparametric flow cytometry. 90/298 (30.2%) AGYW were negative for BV, candidiasis and bacterial STIs. L. crispatus and L. iners were the dominant bacteria in cervicovaginal swabs, and the median vaginal pH was 4.7. AGYW with L. crispatus-dominant microbiota (42.4%) generally had the lowest cytokine concentrations compared to women with more diverse microbiota (34/44 significantly upregulated cytokines). Frequencies of CCR5+CD4+ T-cells co-expressing CD38 and HLA-DR correlated positively with interleukin (IL)-6, TNF-α, GRO-α, macrophage inflammatory protein (MIP)-1α, and IL-9. While endogenous oestrogen had an immune-dampening effect on IL-6, TNF-related apoptosis-inducing ligand (TRAIL) and IL-16, injectable hormone contraceptives (DMPA and Net-EN) were associated with significantly lower endogenous hormone concentrations (p<0.0001 for oestrogen and progesterone) and upregulation of 34/44 cytokines. Since genital inflammation and the presence of activated CD4+ T cells in the genital tract have been implicated in increased HIV risk in South African women, the observed high levels of genital cellular activation and cytokines from AGYW may point towards biological factors increasing HIV risk in this region.

Evidence for cross-protection but not type-replacement over the 11 years after human papillomavirus vaccine introduction

ABSTRACT Examination of cross-protection and type replacement after human papillomavirus (HPV) vaccine introduction is essential to guide vaccination recommendations and policies. The aims of this study were to examine trends in non-vaccine-type HPV: 1) genetically related to vaccine types (to assess for cross-protection) and 2) genetically unrelated to vaccine types (to assess for type replacement), among young women 13–26 years of age during the 11 years after HPV vaccine introduction. Participants were recruited from a hospital-based teen health center and a community health department for four cross-sectional surveillance studies between 2006 and 2017. Participants completed a survey that assessed sociodemographic characteristics and behaviors, and cervicovaginal swabs were collected and tested for 36 HPV genotypes. We determined changes in proportions of non-vaccine-type HPV prevalence and conducted logistic regression to determine the odds of infection across the surveillance studies, propensity-score adjusted to control for selection bias. Analyses were stratified by vaccination status. Among vaccinated women who received only the 4-valent vaccine (n = 1,540), the adjusted prevalence of HPV types genetically related to HPV16 decreased significantly by 45.8% (adjusted odds ratio [AOR] = 0.48, 95% confidence interval [CI] = 0.31–0.74) from 2006–2017, demonstrating evidence of cross-protection. The adjusted prevalence of HPV types genetically related to HPV18 did not change significantly (14.2% decrease, AOR = 0.83, 95% CI = 0.56–1.21). The adjusted prevalence of HPV types genetically unrelated to vaccine types did not change significantly (4.2% increase, AOR = 1.09, CI = 0.80–1.48), demonstrating no evidence of type replacement. Further studies are needed to monitor for cross-protection and possible type replacement after introduction of the 9-valent HPV vaccine.

The rates of mutations associated with macrolide resistance in Mycoplasma genitalium among patients with non-gonococcal sexually transmitted infections in Smolensk and Tula

Objective. In this study, we examined the prevalence of macrolide-resistant M. genitalium in two Russian cities, Smolensk and Tula, between 2013 and 2017. Materials and Methods. DNA’s samples were isolated from urethral and cervicovaginal swabs using primary screening and tested for macrolide resistance-associated mutations by real-time PCR. This technology makes it possible to identify any nucleotide substitutions in the 23S rRNA M. genitalium gene at positions 2058, 2059, 2611 M. genitalium by melting curve analysis after the amplification. Results. According to the study in two cities (Smolensk and Tula) macrolide resistance-associated mutations were found in 3, 65% of isolates (21⁄574). The A2058G transition 23S rRNA MGE was the most common mutation that is associated with macrolide resistance: 5⁄12 (41.6%) – Smolensk, 8⁄9 (88.8%) – Tula. Rare substitutions have been reported at position A2058T 23S rRNA MGE and at position C2611T23S rRNA MGE. All received data is published at the AntiMicrobial Resistance Map (AMRmap) (http://AMRmap.com). Conclusions. According to our study, the frequency of macrolide-resistance mutations in M. genitalium was not more than 4% in two cities of Central Russia during 2013–2017. Despite the relatively low rates of resistance of M. genitalium to macrolides in Smolensk and Tula, our data emphasize the need for epidemiological surveillance of resistance in this pathogen.

Use of Multiplex Polymerase Chain Reaction for Detection of High-Risk Human Papillomavirus Genotypes in Women Attending Routine Cervical Cancer Screening in Harare

Background/Aims: In Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness of these methods can be increased if complemented by a human papillomavirus DNA detection tool since most cervical cancer cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV) genotypes. Moreover, the possibility of multiple-genotype HR-HPV infections warrants the need for HPV detection tools with the capacity to detect both single and multiple infections. The aim of this study was to detect HR-HPV genotypes (HPV 16, 18, 31, 33, 35, 45, 51, 52, 56, and 58), using multiplex polymerase chain reaction (PCR), in stored cervicovaginal swabs from both HIV-positive and HIV-negative women reporting for routine cervical cancer screening. Methodology: Stored cervicovaginal swabs from sexually active women who underwent VIAC at the Parirenyatwa Referral Hospital in Harare, Zimbabwe, between February and April 2015 and had received HIV counselling and testing were genotyped for the selected 10 HR-HPV genotypes using in-house multiplex PCR. The results from the multiplex PCR were compared to those previously obtained when the same samples were HPV genotyped with next-generation sequencing (NGS) on an MiSeq platform (Illumina; USA). Results: A total of 136 women were recruited and all 10 HR-HPV genotypes were detected. Quality control failed in 3 of the 136 swabs during the multiplex PCR reactions. The prevalence of HR-HPV genotypes in the study subjects was 53% (70/133). HIV-infected women were 1.67 times more likely to be infected with HR-HPV than were HIV-negative women (OR 1.67; p = 0.17). Of the 70 HR-HPV-positive cases, 37% (26/70) had multiple HR-HPV infections, and the majority of them were HIV infected. HIV-infected women were 1.86 times more likely to have multiple HR-HPV infections than HIV-negative women (OR 1.86; p = 0.20). Multiplex PCR and NGS had an almost perfect concordance rate in ­HR-HPV detection (κ = 0.960), with only 3 discordant cases (negative with NGS and positive for HPV16 with multiplex PCR). Conclusion: Multiplex PCR can detect HR-HPV genotypes that are common in Zimbabwe and could be used to detect HR-HPV genotypes from women attending cervical cancer screening programs at the Parirenyatwa VIAC clinic in Harare.

Epidemiological evaluation of human papillomavirus genotypes and their associations in multiple infections.

The aim of this study was to determine the frequency of multiple type human papillomavirus (HPV) infections, and whether any types are involved in multiple HPV-type infections (mHPV) more or less frequently than expected. From January 2012 to February 2018, 2848 cervico-vaginal swabs were analysed in the UOC Microbiology and Virology of Policlinico of Bari, Italy. HPV DNA detection was performed using initially nested-polymerase chain reaction (PCR) and subsequently multiplex real-time PCR assay. 1357/2848 samples (47.65%) were HPV DNA positive and 694/1357 (51.14%) showed mHPVs. The median number of mHPVs was 2 (interquartile range: 2-3). HPV-types more frequently detected were 42 (9.97%), 16 (8.92%), 53 (7.23%) and 31 (7.16%). Each detected HPV-type was involved in mHPVs in more than 50% of cases. Statistical analysis showed significant associations for all HPV-types except for 33, 43, 51, 58 and 82 HPV-types. The major number of significant pairwise associations were detected for the types 42 and 70. Only positive associations were detected. Further data are necessary to evaluate the clinical impact of the single combinations.

A Head-to-Head Analytical Comparison of Cobas 4800 HPV, PapilloCheck HPV Screening, and LMNX Genotyping Kit HPV GP for Detection of Human Papillomavirus DNA in Cervical and Cervicovaginal Swabs

High-risk human papillomavirus (hrHPV) infection is a cause of cervical cancer development. The addition of hrHPV testing to cervical cancer screening and monitoring of cervical intraepithelial neoplasia treatment improves the efficacy of screening and treatment, respectively. Self-sampling for hrHPV testing seems a promising tool for increasing patient participation in cervical cancer screening. In this project, 1198 cervical swabs obtained by physicians and 176 cervicovaginal swabs obtained by self-sampling (not collected in parallel) were analyzed for the presence of 14 hrHPV genotypes using three commercially available assays in comparison. HPV DNA was detected in 21.2% of all samples (21% of cervical swabs and 22.7% of cervicovaginal swabs). The cobas 4800 HPV Test was the most sensitive (0.983) and specific (0.992) for hrHPV detection overall. The PapilloCheck HPV-Screening and LMNX Genotyping Kit HPV GP had comparable specificity with that of the cobas (0.989 and 0.955, respectively), but lesser sensitivity (0.897 and 0.909, respectively). In physician-obtained cervical swabs, the cobas showed the highest sensitivity and specificity (0.980 and 0.994, respectively) for hrHPV detection, whereas in cervicovaginal swabs, the cobas had the highest sensitivity (1.00), but the PapilloCheck had the highest specificity (0.993). In conclusion, all of the detection methods evaluated were highly sensitive and specific for hrHPV detection from both clinician-collected cervical swabs andself-sampled cervicovaginal swabs.

Prevalence, incidence and correlates of low risk HPV infection and anogenital warts in a cohort of women living with HIV in Burkina Faso and South Africa.

ObjectiveTo report the prevalence and incidence of low-risk human papillomavirus infection (LR-HPV) and anogenital warts (AGW) among women living with HIV (WLHIV) in Burkina Faso (BF) and South Africa (SA), and to explore HIV-related factors associated with these outcomes.MethodsWe enrolled 1238 WLHIV (BF = 615; SA = 623) aged 25–50 years and followed them at three time points (6, 12 and 16 months) after enrolment. Presence of AGW was assessed during gynaecological examination. Cervico-vaginal swabs for enrolment and month 16 follow-up visits were tested for HPV infection by Inno-LiPA® genotyping. Logistic regression was used to assess risk factors for prevalent infection or AGW. Cox regression was used to assess risk factors for incident AGW.ResultsWomen in SA were more likely than those in BF to have prevalent LR-HPV infection (BF: 27.1% vs. SA: 40.9%; p<0.001) and incident LR-HPV infection (BF: 25.8% vs. SA: 31.6%, p = 0.05). Prevalence of persistent LR-HPV was similar in the two countries (BF: 33.3% vs. SA: 30.4%; p = 0.54), as were prevalence and incidence of AGW (Prevalence: BF: 7.5% vs. SA: 5.7%; p = 0.21; Incidence: BF: 2.47 vs. SA: 2.33 per 100 person-years; p = 0.41). HPV6 was associated with incident AGW (BF: adjusted Hazard Ratio (aHR) = 4.88; 95%CI: 1.36–17.45; SA: aHR = 5.02; 95%CI: 1.40–17.99). Prevalent LR-HPV (BF: adjusted Odds Ratio [aOR = 1.86]; 95%CI: 1.01–3.41; SA: aOR = 1.75; 95%CI: 0.88–3.48); persistent LR-HPV (BF: aOR = 1.92; 95%CI: 0.44–8.44; SA: aOR = 2.81; 95%CI: 1.07–7.41) and prevalent AGW (BF: aOR = 1.53; 95%CI: 0.61–3.87; SA: aOR = 4.11; 95%CI: 1.20–14.10) were each associated with low CD4+ counts (i.e. <200 vs. >500 cells/μL). Duration of ART and HIV plasma viral load were not associated with any LR-HPV infection or AGW outcomes.ConclusionLR-HPV infection and AGW are common in WLHIV in sub-Saharan Africa. Type-specific HPV vaccines and effective ART with immunological reconstitution could reduce the burden of AGW in this population.

Prevalence of Genital Human Papillomavirus Among Sexually Experienced Males and Females Aged 14-59 Years, United States, 2013-2014.

Differences in human papillomavirus (HPV) prevalence among males and females have been reported. Using the 2013-2014 National Health and Nutrition Examination Survey, we evaluated sex differences in prevalence overall and by demographic and sexual behavior characteristics. Self-collected penile and cervicovaginal swabs from participants aged 14-59 were tested for HPV DNA. Prevalences of any HPV and high-risk HPV (HR-HPV) were estimated for sexually experienced males and females. Overall and in models stratified by demographic characteristics and behaviors, prevalence was compared in males and females using prevalence ratios (PR). Overall, males had higher prevalence than females of any HPV (PR, 1.14; 95% confidence interval [CI], 1.03-1.27) and HR-HPV (PR, 1.24; 95% CI, 1.07-1.43). Prevalences were lower among males than females at ages 14-19 and higher at ages 40-49 and 50-59. Sex differences in models stratified by race/ethnicity, poverty, sexual behaviors, and smoking were observed. After adjusting for lifetime sex partners, most sex differences were attenuated, but males had lower prevalences at ages 14-19 and 20-24 and higher HR-HPV prevalence among non-Hispanic blacks. Any HPV and HR-HPV prevalences were significantly higher in males; sex differences varied by age group and race/ethnicity. Lifetime partners explained many of the differences by sex.