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1.
BJOG ; 127(2): 182-192, 2020 01.
Article in English | MEDLINE | ID: mdl-31749298

ABSTRACT

OBJECTIVE: Characterise the vaginal metabolome of cervical HPV-infected and uninfected women. DESIGN: Cross-sectional. SETTING: The Center for Health Behavior Research at the University of Maryland School of Public Health. SAMPLE: Thirty-nine participants, 13 categorised as HPV-negative and 26 as HPV-positive (any genotype; HPV+ ), 14 of whom were positive with at least one high-risk HPV strain (hrHPV). METHOD: Self-collected mid-vaginal swabs were profiled for bacterial composition by 16S rRNA gene amplicon sequencing, metabolites by both gas and liquid chromatography mass spectrometry, and 37 types of HPV DNA. MAIN OUTCOME MEASURES: Metabolite abundances. RESULTS: Vaginal microbiota clustered into Community State Type (CST) I (Lactobacillus crispatus-dominated), CST III (Lactobacillus iners-dominated), and CST IV (low-Lactobacillus, 'molecular-BV'). HPV+ women had higher biogenic amine and phospholipid concentrations compared with HPV- women after adjustment for CST and cigarette smoking. Metabolomic profiles of HPV+ and HPV- women differed in strata of CST. In CST III, there were higher concentrations of biogenic amines and glycogen-related metabolites in HPV+ women than in HPV- women. In CST IV, there were lower concentrations of glutathione, glycogen, and phospholipid-related metabolites in HPV+ participants than in HPV- participants. Across all CSTs, women with hrHPV strains had lower concentrations of amino acids, lipids, and peptides compared with women who had only low-risk HPV (lrHPV). CONCLUSIONS: The vaginal metabolome of HPV+ women differed from HPV- women in terms of several metabolites, including biogenic amines, glutathione, and lipid-related metabolites. If the temporal relation between increased levels of reduced glutathione and oxidised glutathione and HPV incidence/persistence is confirmed in future studies, anti-oxidant therapies may be considered as a non-surgical HPV control intervention. TWEETABLE ABSTRACT: Metabolomics study: Vaginal microenvironment of HPV+ women may be informative for non-surgical interventions.


Subject(s)
Metabolome , Microbiota , Papillomavirus Infections/microbiology , Vagina/microbiology , Adult , Cross-Sectional Studies , Female , High-Throughput Nucleotide Sequencing , Humans , Lactobacillus , Microbiota/genetics , Papillomavirus Infections/genetics , Papillomavirus Infections/virology , RNA, Ribosomal, 16S/genetics , Vagina/virology
2.
Int J STD AIDS ; 23(4): 242-7, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22581946

ABSTRACT

Female sex workers (FSWs) are at high risk of human papillomavirus (HPV) infection. Questionnaires were administered to 200 FSWs aged 18-26 years in Lima, Peru, to gather risk behaviours, and cervical swab samples were collected for Pap smears and HPV DNA testing as part of a longitudinal study. Participants reported a median of 120 clients in the past month, and 99.2% reported using condoms with clients. The prevalence of any HPV in cervical samples was 66.8%; 34 (17.1%) participants had prevalent HPV 16 or 18, and 92 (46.2%) had one or more oncogenic types. Fifteen women had abnormal Pap smears, 13 of which were HPV DNA positive. Fewer years since first sex was associated with oncogenic HPV prevalence in a model adjusted for previous sexually transmitted infection (STI) status and condom use with partners (prevalence ratio = 0.77, 95% confidence interval [CI] = 0.60-0.97). Our data confirm the high rates of HPV transmission among FSWs in Peru, highlighting the need for early and effective strategies to prevent cervical cancer.


Subject(s)
Papillomavirus Infections/epidemiology , Sex Workers , Uterine Cervical Neoplasms/epidemiology , Adolescent , Adult , DNA, Viral/isolation & purification , Female , Humans , Papanicolaou Test , Papillomaviridae/isolation & purification , Papillomavirus Infections/pathology , Peru/epidemiology , Prevalence , Risk Factors , Sexual Behavior/statistics & numerical data , Surveys and Questionnaires , Uterine Cervical Neoplasms/pathology , Vaginal Smears , Young Adult
3.
Int J STD AIDS ; 22(11): 655-8, 2011 Nov.
Article in English | MEDLINE | ID: mdl-22096051

ABSTRACT

Few data exist on oral human papillomavirus (HPV) prevalence in female sex workers (FSWs). Information regarding oral sex practices of 185 Peruvian FSWs, 18-26 years of age, was obtained via survey and compared with HPV testing results of oral rinse samples. Oral HPV prevalence was 14/185 (7.6%); four (28.9%) HPV genotypes were carcinogenic. One hundred and eighty-two participants reported having had oral sex; 95% reported condom use during oral sex with clients and 9.5% with partners. Women who had oral sex more than three times with their partners in the past month were more likely to have oral HPV than women who had oral sex three times or less (P = 0.06). Ten (71.4%) women with oral HPV were HPV-positive at the cervix; conversely 8.3% of women with cervical HPV were HPV-positive in the oral cavity. The prevalence of oral HPV was relatively low, considering the high rates of oral sex practiced by these women.


Subject(s)
Mouth Mucosa/virology , Papillomaviridae/isolation & purification , Papillomavirus Infections/epidemiology , Sex Work , Sexual Behavior/statistics & numerical data , Adolescent , Adult , Female , Genotype , Humans , Papillomaviridae/classification , Papillomaviridae/genetics , Papillomavirus Infections/virology , Peru/epidemiology , Prevalence , Surveys and Questionnaires , Young Adult
4.
J Clin Virol ; 51(1): 44-9, 2011 May.
Article in English | MEDLINE | ID: mdl-21388867

ABSTRACT

INTRODUCTION: While high HPV 16 viral load measured at a single time point is associated with cervical disease outcomes, few studies have assessed changes in HPV 16 viral load on viral clearance. OBJECTIVE: To measure the association between changes in HPV 16 viral load and viral clearance in a cohort of Thai women infected with HPV 16. STUDY DESIGN: Fifty women (n=50) between the ages of 18-35 years enrolled in a prospective cohort study were followed up every three months for two years. Women positive for HPV 16 DNA by multiplex TaqMan assay at two or more study visits were selected for viral load quantitation using a type-specific TaqMan based real-time PCR assay. The strength of the association of change in viral load between two visits and viral clearance at the subsequent visit was assessed using a GEE model for binary outcomes. RESULTS: At study entry, HPV 16 viral load did not vary by infection outcome. A >2 log decline in viral load across two study visits was found to be strongly associated with viral clearance (AOR: 5.5, 95% CI: 1.4-21.3). HPV 16 viral load measured at a single time point was not associated with viral clearance. CONCLUSIONS: These results demonstrate that repeated measurement of HPV 16 viral load may be a useful predictor in determining the outcome of early endpoints of viral infection.


Subject(s)
DNA, Viral/metabolism , Human papillomavirus 16/physiology , Papillomavirus Infections/virology , Uterine Cervical Dysplasia/virology , Uterine Cervical Neoplasms/virology , Viral Load/genetics , Adult , Cervix Uteri/virology , Cohort Studies , DNA, Viral/analysis , DNA, Viral/genetics , Female , Follow-Up Studies , Human papillomavirus 16/genetics , Human papillomavirus 16/immunology , Humans , Kinetics , Papillomavirus Infections/immunology , Polymerase Chain Reaction , Prospective Studies , Thailand , Uterine Cervical Neoplasms/immunology , Viral Load/immunology , Young Adult , Uterine Cervical Dysplasia/immunology
5.
Sex Transm Infect ; 84(4): 306-11, 2008 Aug.
Article in English | MEDLINE | ID: mdl-18385223

ABSTRACT

OBJECTIVE: To investigate self-administered vaginal swabs for assessing prevalence and correlates of carcinogenic human papillomavirus (HPV) infection in rural Rakai, Uganda. METHODS: 1003 sexually experienced women enrolled in a community cohort provided self-administered vaginal swabs collected at annual, home-based surveys. Carcinogenic HPV prevalence, adjusted odds ratios (AOR), 95% confidence intervals (CI) and associated risk factors were determined. RESULTS: Carcinogenic HPV prevalence was 19.2%: 46.6% among HIV positive and 14.8% among HIV negative women (p<0.001). Type-specific prevalence ranged from 2.0% (HPV 16 and 52) to 0.2% (HPV 31). Age-specific HPV prevalence decreased significantly (p<0.001) among HIV negative women; however, the decrease among HIV positive women was not as pronounced (p = 0.1). Factors independently associated with carcinogenic HPV infection were HIV (AOR 4.82, CI 3.10 to 7.53), age (AOR 4.97, 95% CI 2.19 to 11.26 for 15-19 year olds compared to 40+ years), more than two sex partners in the past year (AOR 2.21, CI 1.10 to 4.43) and self-reported herpes zoster, candidiasis or tuberculosis (AOR 4.52, CI 1.01 to 20.31). Married women were less likely to have prevalent carcinogenic HPV (AOR 0.46, CI 0.30 to 0.70). CONCLUSIONS: HPV prevalence and correlates measured using self-administered vaginal swabs were similar to studies that use cervical samples. Thus, self-collection can be used as a substitute for cervical specimens and provide an important tool for research in populations unwilling to undergo pelvic exam.


Subject(s)
Papillomavirus Infections/epidemiology , Adolescent , Adult , DNA, Viral/analysis , Female , Genotype , Humans , Middle Aged , Polymerase Chain Reaction/methods , Prevalence , Risk Factors , Rural Health , Tumor Virus Infections/epidemiology , Uganda/epidemiology
6.
J Infect Dis ; 183(11): 1554-64, 2001 Jun 01.
Article in English | MEDLINE | ID: mdl-11343204

ABSTRACT

This study investigated the association of selected demographic and behavioral characteristics with the detection of low-risk, high-risk, and uncharacterized genital human papillomavirus (HPV) in women attending clinic for routine nonreferral gynecologic health care. Cervical specimens obtained from 3863 women 18-40 years old (mean, 28 years) with no history of high-grade cervical disease were analyzed for 38 HPV types. Overall, HPV prevalence was 39.2%. The prevalence of high-risk, low-risk, and uncharacterized HPV types was 26.7%, 14.7%, and 13.0%, respectively. As expected, the characteristics most strongly associated with overall HPV detection were age and numbers of lifetime and recent sex partners. Low-risk, high-risk, and uncharacterized HPV detection increased with increasing numbers of sex partners. There was a decline in high-risk and low-risk HPV detection with increasing age but little change in uncharacterized HPV detection. These results suggest that the uncharacterized HPV types have a different natural history than either low-risk or high-risk HPV types.


Subject(s)
Genital Diseases, Female/epidemiology , Papillomaviridae/isolation & purification , Papillomavirus Infections/epidemiology , Tumor Virus Infections/epidemiology , Adolescent , Adult , Cervix Uteri/virology , DNA, Viral/analysis , Female , Genital Diseases, Female/virology , Humans , Mass Screening/methods , Odds Ratio , Papillomaviridae/genetics , Papillomavirus Infections/virology , Polymerase Chain Reaction , Risk Factors , Sexual Behavior , Tumor Virus Infections/virology , United States/epidemiology
8.
Cancer Causes Control ; 12(2): 153-61, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11246844

ABSTRACT

OBJECTIVES: Few studies of smoking and cervical carcinoma have addressed the rare cervical adenocarcinomas or used DNA-based tests to control for human papillomavirus (HPV) infection. METHODS: This multicenter case-control study included 124 adenocarcinoma cases, 307 community controls (matched on age, race, and residence to adenocarcinoma cases), and 139 squamous carcinoma cases (matched on age, diagnosis date, clinic, and disease stage to adenocarcinoma cases). Participants completed risk-factor interviews and volunteered cervical samples for PCR-based HPV testing. Polychotomous logistic regression generated adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for both histologic types. RESULTS: Eighteen percent of adenocarcinoma cases, 43% of squamous carcinoma cases, and 22% of controls were current smokers. After control for HPV and other questionnaire data, adenocarcinomas were consistently inversely associated with smoking (e.g. current: OR = 0.6, 95% CI 0.3-1.1; > or = 1 pack per day: OR = 0.7, 95% CI 0.4-1.3), while squamous carcinomas were positively associated with smoking (e.g. current: OR = 1.6, 95% CI 0.9-2.9; > or = 1 pack per day: OR = 1.8, 95% CI 1.0-3.3). Results in analyses restricted to HPV-positive controls were similar. CONCLUSION: Smoking has opposite associations with cervical adenocarcinomas and squamous carcinomas. Although both histologic types are caused by HPV and arise in the cervix, etiologic co-factors for these tumors may differ.


Subject(s)
Adenocarcinoma/epidemiology , Carcinoma, Squamous Cell/epidemiology , Smoking/epidemiology , Uterine Cervical Neoplasms/epidemiology , Adenocarcinoma/diagnosis , Adult , Age Distribution , Aged , Carcinoma, Squamous Cell/diagnosis , Case-Control Studies , Cohort Studies , Comorbidity , Confidence Intervals , Female , Humans , Incidence , Logistic Models , Middle Aged , Multicenter Studies as Topic , Odds Ratio , Risk Factors , Time Factors , United States/epidemiology , Uterine Cervical Neoplasms/diagnosis
9.
Cancer Epidemiol Biomarkers Prev ; 10(2): 95-100, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11219778

ABSTRACT

As human papillomavirus (HPV) becomes accepted as the central cause of cervical cancer, longitudinal studies are shifting focus away from causality to a more detailed investigation of the natural history of HPV infections. These studies commonly require repeated samples for HPV testing over several years, usually collected during a pelvic exam, which is inconvenient to the participants and costly to the study. To alleviate the inconvenience and cost of repeated clinic visits, it has been proposed that women collect cervicovaginal cells themselves, hopefully increasing participation in the natural history studies. We evaluated the technical feasibility of self-collection of cervicovaginal cells using a Dacron swab for HPV DNA detection. We compared the self-collected swab sample and two clinician-administered swab samples (one from the endocervix and another from the ectocervix) from a total of 268 women participating in a case-control study of adenocarcinoma and squamous cell carcinomas of the uterine cervix (111 cases and 157 controls). HPV DNA was detected and genotyped using an L1 consensus PCR assay. The overall agreement between the clinician- and self-collected swabs was excellent [88.1%; kappa = 0.73 (95% confidence interval (CI), 0.61-0.85)]. The correlation was highest between the two clinician-administered swabs [kappa = 0.81 (95% CI, 0.69-0.93)] but was still excellent when comparing either clinician-administered swab to the self-administered sample [kappa = 0.75 (95% CI, 0.63-0.87) and 0.67 (95% CI, 0.55-0.79) for ectocervix and endocervix, respectively]. The type-specific agreement between samples was higher for high-risk, or cancer-associated, HPV genotypes than for low risk, noncancer-associated HPV genotypes when comparing the self-administered swab sample to the clinician-administered swab sample (kappa = 0.78 for high-risk versus 0.66 for low-risk HPV infections, t = -1.45, P = 0.15). The decrease in agreement for low risk types was largely attributable to an increased detection of these types in the self-administered sample (McNemar's chi2 = 6.25, P = 0.01 for clinician- versus self-administered swab comparisons). The agreement did not vary significantly by age, menopausal status, case status, or clinic center. We have demonstrated that a self-collected Dacron swab sample of cervicovaginal cells is a technically feasible alternative to clinician-administered cervical cell collection in natural history studies of HPV and cervical cancer.


Subject(s)
Papillomaviridae/isolation & purification , Papillomavirus Infections/diagnosis , Polymerase Chain Reaction/methods , Tumor Virus Infections/diagnosis , Vaginal Smears/methods , Adolescent , Adult , Age Distribution , Aged , Case-Control Studies , Confidence Intervals , DNA, Viral/analysis , Female , Humans , Middle Aged , Papillomavirus Infections/epidemiology , Patient Participation , Prevalence , Risk Factors , Sampling Studies , Sensitivity and Specificity , Tumor Virus Infections/epidemiology
10.
Gynecol Oncol ; 77(1): 149-54, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10739704

ABSTRACT

INTRODUCTION: Exogenous hormones may influence the development of cervical adenocarcinomas. Incidence rates of adenocarcinomas and use of noncontraceptive hormones have increased since the 1970s, but few studies have investigated this potential relationship. METHODS: We conducted a multicenter case-control study of 124 women with adenocarcinomas, 139 women with squamous cell carcinomas matched on age, diagnosis date, clinic, and stage of disease (in situ or invasive) to adenocarcinoma cases, and 307 healthy community controls who were also matched on age, ethnicity, and residence to adenocarcinoma cases. Participants completed in-person interviews regarding exogenous hormone use before diagnosis and other risk factors and volunteered cervical samples for human papillomavirus (HPV) testing via a PCR-based method. Odds ratios (ORs) with 95% confidence intervals (CIs) estimated relative risks. RESULTS: Only 13 adenocarcinoma cases (10.5%), 7 squamous carcinoma cases (5%), and 20 controls (6.5%) had used noncontraceptive hormones for menopausal symptoms, irregular periods, or disease prevention; most use was short-term, former use. Ever-use was associated with adenocarcinomas (OR = 2.1, 95% CI 0.95-4.6) but not squamous carcinomas (OR = 0.85, 95% CI 0.34-2.1). No trends were seen with duration of use or ages at first use, but unopposed estrogens were positively associated with adenocarcinomas (OR = 2.7). Unopposed estrogens remained associated with adenocarcinomas (OR = 2.0) when analyses were restricted to the HPV-positive controls. Menopausal status was not associated with adenocarcinomas or squamous carcinomas and did not modify the other associations. CONCLUSION: Although small numbers warrant tentative conclusions, exogenous estrogens, especially unopposed estrogens, were positively associated with adenocarcinomas. Noncontraceptive hormones were negatively but weakly associated with squamous carcinomas.


Subject(s)
Adenocarcinoma/etiology , Carcinoma, Squamous Cell/etiology , Hormone Replacement Therapy/adverse effects , Uterine Cervical Neoplasms/etiology , Adult , Aged , Case-Control Studies , Estrogens/adverse effects , Estrogens/therapeutic use , Female , Humans , Menopause , Middle Aged , Risk Assessment
11.
J Clin Microbiol ; 38(1): 357-61, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10618116

ABSTRACT

Genital human papillomaviruses (HPVs) are commonly detected from clinical samples by consensus PCR methods. Two commonly used primer systems, the MY09-MY11 (MY09/11) primers and the GP5+-GP6+ (GP5+/6+) primers, amplify a broad spectrum of HPV genotypes, but with various levels of sensitivity among the HPV types. Analysis of the primer-target sequence homology for the MY09/11 primers showed an association between inefficient amplification of HPV types and the number and position of mismatches, despite accommodation of sequence variation by inclusion of degenerate base sites. The MY09/11 primers were redesigned to increase the sensitivity of amplification across the type spectrum by using the same primer binding regions in the L1 open reading frame. Sequence heterogeneity was accommodated by designing multiple primer sequences that were combined into an upstream pool of 5 oligonucleotides (PGMY11) and a downstream pool of 13 oligonucleotides (PGMY09), thereby avoiding use of degenerate bases that yield irreproducible primer syntheses. The performance of the PGMY09-PGMY11 (PGMY09/11) primer system relative to that of the standard MY09/11 system was evaluated with a set of 262 cervicovaginal lavage specimens. There was a 91.5% overall agreement between the two systems (kappa = 0.83; P < 0.001). The PGMY09/11 system appeared to be significantly more sensitive than the MY09/11 system, detecting an additional 20 HPV-positive specimens, for a prevalence of 62.8% versus a prevalence of 55.1% with the MY09/11 system (McNemar's chi(2) = 17.2; P < 0.001). The proportion of multiple infections detected increased with the PGMY09/11 system (40. 0 versus 33.8% of positive infections). HPV types 26, 35, 42, 45, 52, 54, 55, 59, 66, 73, and MM7 were detected at least 25% more often with the PGMY09/11 system. The PGMY09/11 primer system affords an increase in type-specific amplification sensitivity over that of the standard MY09/11 primer system. This new primer system will be useful in assessing the natural history of HPV infections, particularly when the analysis requires HPV typing.


Subject(s)
Genital Diseases, Female/virology , Papillomaviridae/genetics , Papillomavirus Infections/virology , Polymerase Chain Reaction/methods , Tumor Virus Infections/virology , Base Sequence , Cervix Uteri/virology , Consensus Sequence , DNA Primers , Female , Humans , Male , Papillomaviridae/isolation & purification , Therapeutic Irrigation , Vagina/virology
12.
Cancer Epidemiol Biomarkers Prev ; 8(12): 1079-85, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10613340

ABSTRACT

To assess the hypothesis that oral contraceptives (OCs) increase the risk of cervical adenocarcinomas, we conducted a six-center case-control study of 124 patients with adenocarcinomas, 139 with squamous cell carcinomas, and 307 population controls. Women between the ages of 18 and 69 who were newly diagnosed with cervical adenocarcinomas between 1992 and 1996 were eligible. Healthy female controls and a second case group of incident cervical squamous cell carcinomas were matched to the adenocarcinoma cases. All participants were interviewed regarding OCs, other risk factors for cervical carcinoma, and utilization of cytological screening, and a PCR-based test determined HPV genotype of cervical samples for both case groups and controls. Use of OCs was positively and significantly associated with adenocarcinomas and positively but weakly associated with squamous cell carcinomas. Associations between OCs and invasive adenocarcinomas (n = 91), squamous cell carcinoma in situ (n = 48), and invasive squamous cell carcinomas (n = 91) disappeared after accounting for HPV infection, sexual history, and cytological screening, but a positive association remained between current use of OCs and cervical adenocarcinoma in situ (n = 33). This association persisted after stratification by screening and sexual history and after restriction according to HPV status, but small numbers made it difficult to exclude detection bias, selection bias, or residual confounding by HPV as potential explanations Current OC use was associated with cervical adenocarcinomas in situ, but we saw no other evidence that OCs independently increase the risk of cervical carcinomas.


Subject(s)
Adenocarcinoma/chemically induced , Carcinoma, Squamous Cell/chemically induced , Contraceptives, Oral/adverse effects , Uterine Cervical Neoplasms/chemically induced , Adenocarcinoma/etiology , Adenocarcinoma/pathology , Adolescent , Adult , Aged , Bias , Carcinoma, Squamous Cell/etiology , Carcinoma, Squamous Cell/pathology , Case-Control Studies , Confounding Factors, Epidemiologic , DNA, Neoplasm/analysis , Female , Humans , Mass Screening , Middle Aged , Neoplasm Staging , Papillomaviridae , Papillomavirus Infections/complications , Polymerase Chain Reaction , Risk Factors , Sexual Behavior , Tumor Virus Infections/complications , Uterine Cervical Neoplasms/etiology , Uterine Cervical Neoplasms/pathology
13.
J Clin Microbiol ; 36(10): 3020-7, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9738060

ABSTRACT

Amplification of human papillomavirus (HPV) DNA by L1 consensus primer systems (e.g., MY09/11 or GP5(+)/6(+)) can detect as few as 10 to 100 molecules of HPV targets from a genital sample. However, genotype determination by dot blot hybridization is laborious and requires at least 27 separate hybridizations for substantive HPV-type discrimination. A reverse blot method was developed which employs a biotin-labeled PCR product hybridized to an array of immobilized oligonucleotide probes. By the reverse blot strip analysis, genotype discrimination of multiple HPV types can be accomplished in a single hybridization and wash cycle. Twenty-seven HPV probe mixes, two control probe concentrations, and a single reference line were immobilized to 75- by 6-mm nylon strips. Each individual probe line contained a mixture of two bovine serum albumin-conjugated oligonucleotide probes specific to a unique HPV genotype. The genotype spectrum discriminated on this strip includes the high-risk, or cancer-associated, HPV genotypes 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 55, 56, 58, 59, 68 (ME180), MM4 (W13B), MM7 (P291), and MM9 (P238A) and the low-risk, or non-cancer-associated, genotypes 6, 11, 40, 42, 53, 54, 57, 66, and MM8 (P155). In addition, two concentrations of beta-globin probes allowed for assessment of individual specimen adequacy following amplification. We have evaluated the performance of the strip method relative to that of a previously reported dot blot format (H. M. Bauer et al., p. 132-152, in C. S. Herrington and J. O. D. McGee (ed.), Diagnostic Molecular Pathology: a Practical Approach, (1992), by testing 328 cervical swab samples collected in Digene specimen transport medium (Digene Diagnostics, Silver Spring, Md.). We show excellent agreement between the two detection formats, with 92% concordance for HPV positivity (kappa = 0.78, P < 0.001). Nearly all of the discrepant HPV-positive samples resulted from weak signals and can be attributed to sampling error from specimens with low concentrations (<1 copy/microliter) of HPV DNA. The primary advantage of the strip-based detection system is the ability to rapidly genotype HPVs present in genital samples with high sensitivity and specificity, minimizing the likelihood of misclassification.


Subject(s)
Papillomaviridae/classification , Papillomaviridae/genetics , Papillomavirus Infections/virology , Tumor Virus Infections/virology , Animals , Base Sequence , Biotinylation , Cattle , Cervix Uteri/virology , Consensus Sequence , Female , Genotype , Humans , Oligonucleotide Probes , Papillomaviridae/isolation & purification , Polymerase Chain Reaction/methods , Random Amplified Polymorphic DNA Technique , Sensitivity and Specificity
14.
Genome Res ; 5(1): 79-88, 1995 Aug.
Article in English | MEDLINE | ID: mdl-8717058

ABSTRACT

Recently, several improvements of traditional PCR techniques have facilitated the amplification of significantly longer DNA target sequences. Here we report an improved method for amplification of entire human papillomavirus (HPV) genomes. Using rTth DNA polymerase, XL (Perkin-Elmer, Foster City CA), and the accompanying XL PCR buffer system, we have successfully amplified 8-kb genomes from approximately 10 copies of input reference strain HPV16 DNA. This long PCR (LPCR) method was subsequently used to amplify the entire HPV16 genome from clinical specimens. The fidelity with which the rTth DNA polymerase XL amplified target sequences under our chosen amplification conditions was estimated by partial sequencing of cloned LPCR products generated from cloned reference strain HPV16 genomes. A region spanning the HPV16 E6, E7, and part of the E1 open reading frames (ORFs) was sequenced in 29 clones. A total of 33 nucleotide substitutions were observed in the 23.5 kb sequenced. This corresponds to an error frequency of approximately one error per 700 bases. Finally, LPCR methods were used to amplify entire, novel HPV genomes from clinical specimens. LPCR primer pairs were designed for amplification of seven potentially novel HPV types. Amplicons of approximately 8 kb were generated from five of the seven HPV types targeted. One of the LPCR-generated novel genomes, CP141, was subsequently cloned and a partial sequence was determined.


Subject(s)
Genome, Viral , Mutagenesis , Papillomaviridae/genetics , Polymerase Chain Reaction/methods , Amino Acid Sequence , Base Sequence , Cervix Mucus/virology , Chromosome Mapping , Cloning, Molecular , DNA Primers , DNA Probes, HPV , DNA, Viral/genetics , Female , Humans , Molecular Sequence Data , Reproducibility of Results , Sensitivity and Specificity , Sequence Alignment , Sequence Homology, Amino Acid
15.
J Infect Dis ; 169(2): 235-40, 1994 Feb.
Article in English | MEDLINE | ID: mdl-8106758

ABSTRACT

Determinants of genital human papillomavirus (HPV) persistence in 393 women initially cytologically normal were investigated by testing them for HPV DNA twice over a median interval of 14.9 months. At each visit, interview information was obtained and a cervicovaginal lavage sample was collected for polymerase chain reaction-based HPV testing. Twenty-six percent of the women were HPV-positive at the first sampling. Data on HPV type was available for 86 HPV-positive women (84%); 35 of these women (41%) had persistent type-specific HPV detection. Persistence decreased with time between samplings. Women aged > or = 30 years had a higher percentage of persistence (65%) than those < or = 24 years (32%, P = .02). The percentage of persistence was higher among women infected with HPV types known to be cancer-associated (45%) than among those infected with other types (24%, P = .11). These findings were independent of each other and of timing between samplings. Although based on a prevalent cohort, these results are concordant with previous suggestions that HPV infection is usually transient and that cervical cancer may arise from within the subset of women with persistent HPV infection.


Subject(s)
Papillomavirus Infections/virology , Tumor Virus Infections/virology , Adolescent , Adult , Age Factors , Aged , Base Sequence , Cervix Uteri/virology , DNA Primers/chemistry , DNA Probes, HPV , Female , Humans , Middle Aged , Molecular Sequence Data , Oligonucleotide Probes/chemistry , Papillomaviridae , Papillomavirus Infections/pathology , Time Factors , Tumor Virus Infections/pathology
16.
J Clin Microbiol ; 31(4): 1003-6, 1993 Apr.
Article in English | MEDLINE | ID: mdl-8385147

ABSTRACT

The detection and classification of human papillomavirus (HPV) by a consensus primer polymerase chain reaction (PCR) technique were compared with detection and classification by dot filter hybridization (DFH) and Southern transfer hybridization (STH). PCR detected HPV in 87% of specimens; the detection rates for DFH and STH were 51% and 49%, respectively. The specific HPV types detected by STH were also detected by PCR in 90% of specimens. However, 75% of the samples positive for unclassified HPV by STH were typed by PCR. PCR results were reproducible, as assessed by repeat analysis (96% agreement), by analysis of paired same-day specimens (89% agreement), and by interlaboratory analysis (88% agreement). PCR is a sensitive, specific, and reproducible test for HPV detection and classification in clinical and epidemiologic studies.


Subject(s)
Anus Diseases/diagnosis , Nucleic Acid Hybridization/methods , Papillomaviridae/isolation & purification , Polymerase Chain Reaction , Tumor Virus Infections/diagnosis , DNA, Viral/analysis , Humans , Male , Papillomaviridae/classification , Papillomaviridae/genetics , Reproducibility of Results , Sensitivity and Specificity
18.
J Urol ; 148(6): 1822-6, 1992 Dec.
Article in English | MEDLINE | ID: mdl-1279224

ABSTRACT

Human papillomavirus is associated with a variety of anogenital lesions, including genital warts, precancers and cancers. In male patients human papillomavirus has been identified in proliferative lesions ranging from penile and urethral warts to penile and prostatic cancers. We examined the association of human papillomavirus deoxyribonucleic acid (DNA) in 84 prostate tissue specimens. Specimens were selected from radical prostatectomy, transurethral resection or transrectal biopsy procedures. A total of 60 formalin-fixed, paraffin-embedded tissues (24 prostate cancer specimens, 16 benign prostatic hyperplasia specimens and 20 normal specimens) was examined by polymerase chain reaction and in situ hybridization. Also, 24 gelatin-embedded frozen prostate cancer specimens were examined for human papillomavirus DNA by polymerase chain reaction. Of the specimens 69 were deemed adequate for polymerase chain reaction analysis, whereas all 60 paraffin-embedded tissues were sufficient for in situ hybridization. Human papillomavirus DNA was detected in 2 normal tissues and 6 prostate cancers using polymerase chain reaction. None of the benign prostatic hyperplasia specimens was positive for human papillomavirus. Human papillomavirus typing results indicated that virus type 16 was present in each of the 8 positive specimens. Confirmation of the presence of human papillomavirus was obtained for 1 of the prostate cancers by nonisotopic in situ hybridization with biotinylated human papillomavirus genomic probes. The low prevalence of human papillomavirus in this study population does not strongly support an etiological role for the virus in prostate cancer.


Subject(s)
DNA Probes, HPV , In Situ Hybridization , Papillomaviridae/isolation & purification , Polymerase Chain Reaction , Prostatic Hyperplasia/microbiology , Prostatic Neoplasms/microbiology , Adult , Aged , Aged, 80 and over , Humans , Male , Middle Aged , Papillomaviridae/genetics
19.
Obstet Gynecol ; 79(5 ( Pt 1)): 649-56, 1992 May.
Article in English | MEDLINE | ID: mdl-1314358

ABSTRACT

Conventional wisdom has long held that human papillomavirus (HPV) prevalence is increased in pregnancy. We examined cervical swabs of 375 women for HPV DNA using polymerase chain reaction (PCR) and ViraPap with an expanded probe range. Of this population, 115 were pregnant (42 in the first trimester, 46 in the second, and 27 in the third) and 100 were postpartum. The control population consisted of 160 women who were otherwise similar to the pregnant population but were not pregnant or postpartum. Crude associations were examined between HPV prevalence at defined high, low, and overall levels and the pregnancy status. Multivariate analysis indicated no statistically significant association between the prevalence at any level of infection and pregnancy status. As expected, associations were found between measures of HPV prevalence and both Papanicolaou smear results and warts seen on examination. No association was found for race, smoking behavior, or number of sexual partners and HPV prevalence. The estimated risk of an HPV infection decreased as the age of the women increased. We conclude that a significant relationship between pregnancy and HPV prevalence has yet to be established.


Subject(s)
Papillomaviridae/isolation & purification , Pregnancy Complications, Infectious/epidemiology , Tumor Virus Infections/epidemiology , Adult , Condylomata Acuminata/diagnosis , Condylomata Acuminata/epidemiology , Condylomata Acuminata/microbiology , Female , Genital Neoplasms, Female/diagnosis , Genital Neoplasms, Female/epidemiology , Genital Neoplasms, Female/microbiology , Humans , Polymerase Chain Reaction , Pregnancy , Pregnancy Complications, Infectious/diagnosis , Pregnancy Complications, Neoplastic/diagnosis , Pregnancy Complications, Neoplastic/epidemiology , Pregnancy Complications, Neoplastic/microbiology , Prevalence , Risk Factors , Tumor Virus Infections/complications , Tumor Virus Infections/diagnosis , Uterine Cervical Dysplasia/complications , Uterine Cervical Dysplasia/microbiology , Vagina/microbiology
20.
IARC Sci Publ ; (119): 121-33, 1992.
Article in English | MEDLINE | ID: mdl-1330905

ABSTRACT

The unprecedented sensitivity of the polymerase chain reaction (PCR) DNA amplification technology provides numerous advantages for the detection of human papillomavirus (HPV) DNA. In this chapter, the principles of the method and its advantages are discussed; disadvantages of the technology and proposed solutions are also presented. To accommodate the complexity of the many distinct HPV types, both type-specific and consensus PCR-based detection methods have evolved. Consensus primer systems, which afford the amplification of a broad spectrum of HPV types, have proven particularly useful in epidemiological studies and in novel virus identification. Since the introduction of PCR into the HPV field, there have been rapid advances and improvements PCR technology. However, standardization of PCR methods for HPV detection is essential for research to fully benefit from the technology. To that end, specific guidelines regarding experimental controls, assessment of analytic sensitivity and data interpretation are outlined here.


Subject(s)
DNA, Viral/analysis , Papillomaviridae/genetics , Polymerase Chain Reaction/methods , Humans , Papillomaviridae/classification
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