Sensitivity Of Human Papillomavirus Detection Research Articles

HPV detection in oral mucosa samples in pediatric patients

ABSTRACT INTRODUCTION: The human papillomavirus (HPV) detection favors treatments for patients with clinical manifestations and limits future consequences for those with asymptomatic infections. OBJECTIVES: Therefore, the present study aimed to evaluate the sensitivity of polymerase chain reaction (PCR) for HPV detection from oral mucosa samples, of asymptomatic patients and patients with clinical manifestations of laryngeal papillomatosis. MATERIAL AND METHODS: A total of 49 pediatric patient samples were obtained by exfoliation of the oral mucosa with a sterile brush. The deoxyribonucleic acid (DNA) samples was extracted and used for HPV detection, using GP5 and GP6 oligonucleotides, by conventional PCR and qPCR reactions. RESULTS: Among the 49 samples, eight were from patients clinically diagnosed with laryngeal papillomatosis, but in both conventional PCR and qPCR technic, only one sample had presented positivity. DISCUSSION: These results suggest that the sample type, the methodology used to collect, the extraction methodology used, the anatomical location of the lesion and the oligonucleotides used; all factors strongly influence the sensitivity of HPV detection by PCR methodology. CONCLUSION: Thus, more studies are needed to better determine the sample collection, and the processing techniques present more reproducibility on PCR detection.

Analytical performance of HPV assays on vaginal self-collected vs practitioner-collected cervical samples: the SCoPE study

BackgroundIn the last decade, human papillomavirus (HPV) testing has been evaluated extensively for cervical screening, with studies finding increased sensitivity compared to cytology. Another advantage of HPV based-screening is the ability to test vaginal samples that can be collected by women themselves. Self-collection has the potential to extend cervical screening coverage by increasing participation rates, particularly among women who are under-screened or have never screened. This could have a significant impact on cervical cancer prevention, as the majority of invasive cervical cancer cases occur among under-screened women. Both the Netherlands and Australia have transitioned their national programs from cytology to HPV as the primary screening test and both countries include a pathway for self-collection. ObjectivesWe evaluated the relative sensitivity for HPV detection of self-collection compared with practitioner-collected cervical specimens in the context of the Australian National Cervical Screening Program (NCSP). Study Design303 women aged ≥18 years attending a single tertiary referral centre took their own sample using a flocked-swab, and then had a practitioner-collected sample taken at colposcopy. All samples were tested at a single laboratory on the six PCR-based HPV assays which can be utilised in the NCSP; Roche cobas 4800 and cobas, Abbott RealTime, BD Onclarity, Cepheid Xpert, and Seegene Anyplex. ResultsHPV16/18 results had high observed agreement between self- and practitioner-collected samples on all assays (range: 0.94-0.99), with good agreement for non-HPV16/18 oncogenic HPV types (range: 0.64-0.73). ConclusionsSelf-collection for HPV-based cervical screening shows good concordance and relative sensitivity when compared to practitionercollected samples across assays in the NCSP.

The Utility of Urine-Based Sampling for Cervical Cancer Screening in Low-Resource Settings.

Background:WHO has recommended Visual Inspection with Acetic acid (VIA) or Human Papillomavirus (HPV) DNA testing if feasible, for cervical cancer screening in low income countries. However, the number of women undergoing screening is very low as a result of limited information, inadequate infrastructure and invasive nature of sampling. Methods:A cross sectional study was carried out comparing HPV DNA detection by Polymerase Chain Reaction (PCR) in paired cervical and urine samples procured from histologically confirmed cervical cancer cases. Results:Amongst the samples collected from 114 cervical cancer cases, HPV DNA was tested positive in cervical samples of 89 (78.1%) and urine samples of 55 (48.2%) patients. The agreement between the two sampling methods was 66.7% and the kappa value was 0.35 indicating a fair agreement. The sensitivity of HPV detection using urine samples was 59.6% (95% confidence interval 49.16%-69.15%) and the specificity was 92% (95% confidence interval 75.0%-97.8%). Conclusion:Even though not acceptable as an HPV DNA screening tool due to low sensitivity, the urine sampling method is inexpensive and more socially acceptable for large epidemiological surveys in developing countries to estimate the burden.

High‐risk human papillomavirus detection in oropharyngeal cancers: Comparison of saliva sampling methods

Accumulating evidence has suggested the utility of salivary oral rinse as a diagnostic fluid to detect oral human papillomavirus (HPV) DNA, but there are many methods for collecting saliva. Salivary oral rinse and unstimulated whole mouth saliva samples were collected from 45 oropharyngeal cancer (OPC) patients. We show a positive correlation of HPV-16 E2 (r = 0.95, P < 0.0001) and E6/7 (r = 0.93, P < 0.0001) relative copy number as well as HPV genotypes in both sample methods. There was a significant correlation between the two sample methods in the ratio of HPV16 E2 to E6/7 DNA (r = 0.46, P < 0.01). Consistent with previous studies, a mixed HPV-16 form (episomal and integrated) was commonly found in both saliva and tumor samples. Detection of HPV in saliva samples collected by either method yielded comparable results, and showed good sensitivity for detection of HPV derived from OPC.

Validation of a new HPV self-sampling device for cervical cancer screening: The Cervical and Self-Sample In Screening (CASSIS) study

ObjectiveWe compared the self-sampling performance of the newly designed HerSwab™ device with a physician-collected cervical sample and another self-sample using the cobas® PCR Female swab for the detection of cervical intraepithelial neoplasia (CIN) and cancer. MethodsWomen referred for colposcopy at McGill University affiliated hospital clinics collected two consecutive self-samples, one with HerSwab™ and one with cobas® swab, after receiving instructions. The order of sampling was randomized. The colposcopist then collected a cervical sample and conducted a colposcopic examination. Samples were tested for human papillomavirus (HPV) DNA. Sensitivity and specificity to detect CIN2+ and respective 95% confidence intervals (CI) were calculated to compare sampling approaches. The HPV testing agreement between samples was measured using the Kappa statistic. ResultsOf 1217 women enrolled, 1076 had complete results for HPV and cytology; 148 (13.8%) had CIN1, 147 (13.7%) had CIN2/3, and 5 (0.5%) had cancer. There was very good agreement between methods for HPV detection (HerSwab™ versus physician: kappa=0.84; cobas® swabs versus physician: kappa=0.81; HerSwab™ versus cobas® swabs: kappa=0.87). The sensitivity of HPV detection for CIN2+ was 87.6% (95%CI: 79.8–93.2) with self-sampling using HerSwab™, 88.6% (95%CI: 80.9–94.0) with self-sampling using the cobas® swab, and 92.4% (95%CI: 85.5–96.7) with physician sampling. Corresponding estimates of specificity were 58.1% (95%CI: 54.1–62.1), 55.0% (95%CI: 50.9–59.0) and 58.7% (95%CI: 54.6–62.6). Cytology (ASC-US or more severe) done on the physician-collected specimen was 80.2% (95%CI: 70.8–87.6) sensitive and 61.4% (95%CI: 57.2–65.5) specific for CIN2+. ConclusionsThe HerSwab™ had good agreement with physician sampling in detecting HPV, and adequate performance in detecting high-grade lesions among women referred to colposcopy for abnormal cytology.

Vaginal and Urine Self-sampling Compared to Cervical Sampling for HPV-testing with the Cobas 4800 HPV Test.

To compare human papillomavirus (HPV) DNA detection in self-collected vaginal and urine samples with clinician-taken cervical samples in relation to histology. Self-collected vaginal, urine and clinician-taken cervical samples were analyzed from 218 women with the Cobas 4800 HPV test (Roche Molecular Diagnostics). The sensitivity for detection of HPV in the vaginal self-sampling test was 96.4% and in urine was 83.9% relative to detection by clinician-taken cervical sample. The vaginal self-sampling and the clinician-taken HPV tests had the same sensitivity of 92.8% (95% confidence interval=86.3-96.8%) and specificity for detection of high-grade squamous intraepithelial lesion (HSIL) and adenocarcinoma in situ (AIS). Detection in urine samples had a sensitivity of 76.3% (95% confidence interval=67.9-84.2%) for HSIL/AIS. The Cobas 4800 HPV test detects high-grade pre-cancerous cervical lesions in self-collected vaginal samples with the same high sensitivity as in clinician-taken cervical samples.

Human Papilloma Virus Detection by INNOLiPA HPV in Prostate Tissue from Men of Northeast Mexico

Background:Prostatic adenocarcinoma by Prosate cancer (PCa) is the most prevalent cancer and the second cause of cancer-related death among men in the Western world. Human papilloma virus (HPV) may be considered as a preventable risk factor. In this study, we assessed the frequencies of HPV infection in prostatic adenocarcinoma and benign prostatic hyperplasia (BPH) cases in Northeast Mexico.Materials and Methods:A total of 87 paraffin-embedded blocks (from 25 and 62 patients with definite diagnoses of BPH and adenocarcinoma, respectively) were selected and subjected to INNOLiPA HPV Genotyping to detect 28 high- and low-risk HPV types. The rates of infection were compared in the two studied groups.Results:INNOLiPA HPV demonstrated great sensitivity for HPV detection on paraffin-embedded tissue. Global prevalence was 14.9% (13/87). HPV infection was positive in 19.4% (12/62) of patients with adenocarcinoma and 4.0% (1/25) of patients with BPH. HPV-11, which is considered to be low risk, was more prevalent. Interestingly, one patient with BPH and six with prostate cancer showed examples considered to be high risk (HPV-18, -51, -52, and -66).Conclusion:A higher rate of HPV infection among Mexican patients with prostatic carcinoma than among those with BPH was observed. HPV infections may thus contribute to the risk of prostate cancer. Further studies are required to elucidate any roles of HPV infection in prostate disease in Mexico and the effect of prevention and treatment of HPV infection on prostatic adenocarcinoma.

Analytical performance of Cervista ® HPV 16/18 genotyping test for cervical cytology samples

Background Human papillomavirus (HPV) types 16 and 18 are the 2 most frequent types associated with cervical cancer. Identifying their presence or absence in cervical samples may assist in triaging women for subsequent management. The Cervista ® HPV 16/18 genotyping test specifically detects the presence of HPV 16 and 18 in ThinPrep ® cervical specimens. Objectives The objective was to establish the analytical performance of the CERVISTA HPV 16/18 genotyping test. Study design These studies were performed in support of a regulatory submission to the US Food and Drug Administration. Here we report the analytical sensitivity (limit of detection), accuracy compared to consensus L1 gene PCR/bi-directional sequencing, precision, reproducibility, and cross-reactivity (specificity) of the genotyping test. Results Analytical sensitivity for detection of HPV 16 and 18 ranged between 625 and 1250 copies/reaction for both types. When compared to PCR/sequencing for women with atypical squamous cells of undetermined significance cytology, the positive percent agreement was 94.1% (95% confidence interval [CI], 89.8–96.7) and the negative percent agreement was 85.7% (95% CI, 82.4–88.4). The test demonstrated high within-laboratory and inter-operator precision. Reproducibility within sites and between 3 testing sites resulted in 100% agreement with expected results (150 positive, 90 negative results). The genotyping test did not exhibit cross-reactivity to DNA from common low-risk HPV types and other microorganisms found in the human female reproductive tract. Conclusions These analytical performance data support the use of CERVISTA HPV 16/18 genotyping test for the detection and differentiation of HPV 16 and 18 in ThinPrep cervical cytology specimens.

Quality assurance of genotyping array for detection and typing of human papillomavirus

The aim of this study was to evaluate the sensitivity, specificity, reliability and reproducibility of the EasyChip ® HPV blot for human papillomavirus (HPV) genotyping. Type-specific sensitivity and specificity for 39 types of HPV (HPV 6, 11, 16, 18, 26, 31, 32, 33, 35, 37, 39, 42, 43, 44, 45, 51, 52, 53, 54, 55, 56, 58, 59, 61, 62, 66, 67, 68, 69, 70, 72, 74, 82, CP8061, CP8304, L1AE5, MM4, MM7 and MM8) were examined. The operating environment, reliability, reproducibility and blot interpretation were assessed by a quality assurance system. Each batch experiment contained samples from 89 cervical specimens and 7 extrinsic controls. Caski, HeLa and Jurkat cells, male human blood cell DNA and sterile water were used to assess reliability. Furthermore, pairs of sibling controls were used to assess reproducibility. The overall sensitivity of HPV detection was 1–50 copies of HPV genome equivalent. There was no cross-reactivity with amplicons of other HPV genotypes. One hundred batch experiments demonstrated that the reliability was excellent. The intra-batch and inter-batch reproducibility was 98 and 97%, respectively. It was concluded that the EasyChip ® HPV blot is a highly sensitive, reliable and reproducible tool for detection and identification of HPV genotypes.

Prevalência de achados sugestivos de papilomavírus humano (HPV) em biópsias de carcinoma espinocelular de cavidade oral e orofaringe: estudo preliminar

O papilomavírus humano (HPV) é universalmente aceito como agente causal do câncer de colo uterino e, recentemente, vem se especulando sobre sua possível relação com câncer oral e de orofaringe. O carcinoma espinocelular (CEC) oral representa 90% de todos os tumores malignos que afetam a cavidade bucal. Estudos sobre a prevalência de HPV em pacientes com CEC variam de 0 a 100%. O efeito citopático viral mais conhecido é a coilocitose, considerado "critério maior" na infecção pelo HPV do ponto de vista histopatológico. OBJETIVO: O objetivo deste estudo foi verificar a prevalência de achados sugestivos de HPV - coilocitose - em CEC oral e de orofaringe. FORMA DE ESTUDO: coorte transversal. MATERIAL E MÉTODO: Foram examinadas no microscópio 20 lâminas com o diagnóstico de CEC de cavidade oral ou orofaringe sendo que em 15 delas foi encontrada coilocitose, correspondendo a 75%. RESULTADO: Apesar de termos conhecimento que o método com maior sensibilidade atual para pesquisa de HPV ser a reação de polimerase em cadeia (PCR), iniciamos esta pesquisa com a investigação de coilocitose, o que é muito sugestivo de infecção por HPV. CONCLUSÃO: O estudo em questão trata-se de um projeto-piloto pois será dada continuidade a esta pesquisa através da realização de PCR a fim de confirmar a alta prevalência de infecção por HPV em CEC oral e de orofaringe.

Prevalence of histological findings of human papillomavirus (HPV) in oral and oropharyngeal squamous cell carcinoma biopsies: preliminary study

SummaryHuman papillomavirus (HPV) is considered to be an etiologic agent of cervical cancer and, recently its relation to oral and oropharyngeal cancer has also been investigated. Oral squamous cell carcinoma (SCC) represents 90% of all malignant tumors that affect the oral cavity. The prevalence of HPV in patients with SCC ranges from 0 to 100%. The most known viral cytopathic effect is koilocytosis, considered to be a major characteristic of HPV infection. Aim: The aim of this study was to verify the prevalence of some peculiar characteristics of HPV - koilocytosis - in oral and oropharyngeal SCC. Study design: transversal cohort. Material and method: Twenty slides with oral and/or oropharyngeal SCC were examined under microscopy. Results: in 15 of them, koilocytosis was found, amounting to 75%. Although we know that polymerase chain reaction (PCR) is the method with the best sensitivity for HPV detection, we began this research looking for koilocytosis, which is highly suggestive of HPV infection. Conclusion: This study is a trial project and we will continue this research with PCR measures to confirm this high prevalence of HPV infection in oral and oropharyngeal SCC.

Signal-Amplified Colorimetric In Situ Hybridization for Assessment of Human Papillomavirus Infection in Cervical Lesions

Detection and typing of human papillomavirus (HPV) infection may have a major impact in cervical-screening and follow-up. In this study various commercially available techniques for the detection of HPV were evaluated. HPV-status was determined in 86 samples of cervical cancer by PCR and direct sequencing, catalyzed signal amplified colorimetric DNA in situ hybridization (CSAC- ISH) (GenPoint system, DAKO), immunohistochemistry (IHC) and in 12 selected cases also by conventional, non-amplified ISH. Twenty-one samples of cervical intraepithelial neoplasias grade III (CIN III) were investigated by CSAC-ISH, conventional ISH and by IHC, in corresponding PAP smears HPV-detection and typing was performed by CSAC-ISH and Hybrid Capture test II (HC). In additional 20 PAP smears HPV typing was performed using HC and a novel immunocytochemical system for HPV detection and-typing. CSAC-ISH showed good correlation with PCR analysis in cervical cancers: In 87% of PCR positive cases, HPV infection was also detected by CSAC- ISH (66/76). HPV 16 was detected in 75% of PCR-positive cases (44/59), HPV 18 in 71% of PCR positive cases (5/7). CSAC-ISH detected HPV 31 in only 29% of PCR positive cases (2/7), and HPV 33 in 64% of PCR-positive cases (23/36). Nevertheless, CSAC-ISH- false negative cases for HPV 31 or 33 were nearly always combined infections with other HPV types, which were detectable by CSAC-ISH in most cases. CSAC-ISH revealed HPV infection in 20 of 21 HC-positive cervical smears, while in corresponding biopsies (CIN III) CSAC-ISH detected 100% of HPV infections. Conventional, non-amplified ISH showed significantly lower sensitivity compared with CSAC-ISH, and immunocyto- and -histochemistry were of very low sensitivity for detection of HPV. CSAC-ISH is an easy-to-handle method for detection and typing of cervical HPV infection, and shows sufficient sensitivity for clinical practice.

Polymerase chain reaction detection of human papillomavirus: quantitation may improve clinical utility.

A case-control study compared detection by polymerase chain reaction (PCR) specific for human papillomavirus (HPV) type 16 with restriction enzyme analysis and Southern blot hybridization detection of HPV type 16. Cervicovaginal lavage samples from 64 women with histopathologic evidence of a cervical squamous intraepithelial lesion and 55 samples from cytologically healthy women were studied. Several methods of PCR product analysis, including radioactive and nonradioactive probing, were compared. The sensitivity of HPV detection by PCR when the amplified DNA fragment was visualized on a gel was equivalent to those of detection by restriction enzyme and Southern blot analyses. Hybridization of the PCR product with radioactively or nonradioactively labeled oligonucleotide probes increased the sensitivity of HPV detection by 100-fold. However, an increase in the sensitivity of the assay preferentially identified low levels of the virus in cytologically healthy women. Therefore, the value of HPV detection in identifying women with cervical neoplastic disease was greater, and the odds ratio for the presence of a cervical squamous intraepithelial lesion was higher when the less sensitive modalities were used. These results suggest that quantitation of HPV by PCR may maximize the clinical significance of a positive test result. Further studies will be needed to determine the optimal level of virus detection which has the highest positive predictive value of clinical disease.