Consistency evaluation of Liferiver, Yaneng, Darui, and the Cobas 4800 test for high-risk human papillomavirus screening.

BACKGROUND: In recent years, several high-risk human papillomavirus (HR-HPV) tests have been developed. The assay capabilities need to be systematically reviewed. Here, we compared the clinical sample performance of three novel HR-HPV assays (Liferiver, Yaneng, and Darui) based on different platforms with the widely adopted cobas4800 test. METHODS: A total of 346 cervical swabs from women who were screened for cervical cancer were analyzed for the presence of 14 HR-HPV genotypes. The distinction between the four assays was investigated by the genotyping and direct sequencing. RESULTS: The positive rates of the four assays ranged from 61.56% to 64.16%. The overall concordance was 88.15%. The Yaneng assays displayed the best sensitivity (100%) and specificity (98.43%). The sensitivity (98.17%) and specificity (98.43%) of the Darui assay were superior to those of the cobas4800 test (97.72% and 93.70%, respectively). The Liferiver assay displayed comparable sensitivity with the cobas4800 test (95.89% and 97.72%, respectively). The specificity of the cobas4800 was lower than that of the Liferiver assay (93.70% vs. 97.64%). CONCLUSIONS: The three novel HR-HPV assays displayed good agreement with the cobas4800 test. The analytical performance of all four fulfilled the requirements of sensitivity and specificity for HR-HPV detection.

Quality assurance of human papillomavirus (HPV) testing in the implementation of HPV primary screening in Norway: an inter-laboratory reproducibility study.

BACKGROUND: Human papillomavirus (HPV) testing as primary screening for cervical cancer is currently being implemented in Norway in a randomized controlled fashion, involving three laboratories. As part of the quality assurance programme of the implementation, an evaluation of the inter-laboratory reproducibility of the HPV test was initiated, to ensure satisfactory HPV test reliability in all three laboratories. METHODS: The HPV test used is the cobas 4800 HPV Test, detecting 14 high-risk types with individual HPV genotype results for HPV16 and HPV18. In addition to the three laboratories involved in the implementation, the Norwegian HPV reference laboratory was included as a fourth comparative laboratory. A stratified sample of 500 cervical liquid based cytology (LBC) samples was used in the evaluation, with an aim towards a high-risk HPV positivity of ~25%. Samples were collected at one laboratory, anonymized, aliquoted, and distributed to the other laboratories. RESULTS: Comparison of the test results of all four laboratories revealed a 95.6% agreement, an 86.3% positive agreement and a kappa value of 0.94 (95% CI 0.92-0.97). For negative cytology specimens, there was a 95.8% overall agreement, a 67.4% positive agreement, and a kappa value of 0.88 (95% CI 0.80-0.93). For abnormal cytology specimens, there was a 95.8% overall agreement, a 95.5% positive agreement, and a kappa value of 0.86 (95% CI 0.71-0.97). CONCLUSIONS: The study showed a high inter-laboratory reproducibility of HPV testing, implying satisfactory user performance and reliability in the laboratories involved in the implementation project. This is important knowledge and we recommend similar studies always to be performed prior to the introduction of new screening routines.

Effect of glacial acetic acid treatment of cervical ThinPrep specimens on HPV DNA detection with the cobas 4800 HPV test.

BACKGROUND: Cytology laboratories in the UK routinely treat unsatisfactory cervical liquid-based cytology (LBC) specimens with glacial acetic acid (GAA) to reduce the unsatisfactory rate. However, there is limited published data on the effect of GAA reprocessing on the molecular detection of human papillomavirus (HPV). The aim of this study was to assess the impact of GAA treatment of cervical ThinPrep(®) samples on HPV detection with the cobas(®) 4800 HPV Test (Roche Molecular Systems, Pleasanton, CA, USA). METHODS: Residual ThinPrep samples (n = 121) were selected to provide a range of typical cytology results and enrich the study samples for HPV positivity. Specimens were equally split into two fractions: one part treated with 10% GAA and the other part left untreated. All samples were HPV tested using the cobas 4800 HPV Test, which simultaneously detects a total of 14 high-risk HPV (hrHPV) genotypes and individually identifies HPV16 and HPV18. The HPV positive/negative status of tested samples determined the level of agreement between treated and untreated fractions; one sample failed owing to detection of a clot by the instrument during pipetting, leaving 120 samples in the study. Statistical analysis was performed using an unweighted kappa. RESULTS: Analysis of overall HPV positivity showed 97.5% (117/120) agreement between the treated and untreated fractions with a kappa value of 0.95. There were 63/65 (96.9%) concordant HPV positive and 54/55 (98.2%) concordant HPV negative results. In addition to the three discordant results for overall HPV positivity, there were three HPV type-specific discrepancies giving a total of 114/120 concordant HPV results (95% agreement). CONCLUSIONS: Glacial acetic acid (GAA) treatment of cervical ThinPrep specimens does not have significant adverse affects on HPV detection with the cobas 4800 HPV Test.

High-risk Human Papillomavirus Testing in Young Japanese Women with Atypical Squamous Cells of Undetermined Significance.

INTRODUCTION: The mortality due to uterine cervical cancer has been gradually increasing in women under 40 years of age (U40) in Japan. We investigated the effect of high-risk human papillomavirus (HR-HPV) on U40 subjects without any overt cytological abnormalities. MATERIALS AND METHODS: We retrospectively examined the clinical data, including the findings of a cobas 4800 HPV test that was approved in Japan in 2013 to triage women with atypical squamous cells of undetermined significance (ASC-US) and a histological examination in 589 Japanese women. RESULTS: The overall prevalence rate of HR-HPV was 34.5%. Biopsy-confirmed cervical intraepithelial neoplasia (CIN) 2, or worse (CIN2+) was identified in 45.1% (23/51) of HR-HPV-positive women with ASC-US, who underwent colposcopy immediately. The mean period from the HPV test to the diagnosis of CIN2+ was 3.7 months. CIN2+ was more common (69.6%) in U40 patients. The rates of single or multiple infections of HPV-16, HPV-18, and 12 other HR-HPV (31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) in CIN2+ U40 patients were 31.3%, 0%, and 81.3%, respectively. The relative risk for CIN 2+ among U40 women with HPV-16 was not significantly different from that of the patients with infection of any of the 12 other HR-HPVs. CONCLUSION: The results of this study suggest that the 12 other HR-HPVs have a potential to generate high-grade cervical lesions among young women, and the examination rate of colposcopy should be increased.

Prevalence of high-risk human papillomavirus by cobas 4800 HPV test in urban Peru.

BACKGROUND: Molecular tests allow the detection of high-risk human papillomavirus in cervical samples, playing an important role in the prevention of cervical cancer. OBJECTIVES: We performed a study to determine the prevalence of HPV 16, HPV 18 and other high-risk human papillomavirus (pool 12 genotypes) in Peruvian females from diverse urban areas using the cobas 4800 HPV test. METHODS: Routine cervical samples collected in our laboratory were analyzed by cobas 4800 HPV test. RESULTS: A total of 2247 samples from female patients aged 17-79 years were tested. high-risk human papillomavirus was positive in 775 (34.49%) samples. Of these, 641 (82.71%) were single infections and 134 (17.29%) were multiple infections. The positivity rates for HPV 16, HPV 18, and other high-risk human papillomavirus were 10.77%, 2.0%, and 28.08%, respectively. In multiple high-risk human papillomavirus infections, the concomitance of HPV 16 and other high-risk human papillomavirus was more prevalent (13.42%). CONCLUSION: Our study showed high prevalence of high-risk human papillomavirus in urban Peru, mainly among young women. In both single and multiple infections other high-risk human papillomavirus were more prevalent than HPV 16 and HPV 18, which might influence vaccine impact in our country. Furthermore, the cobas 4800 HPV test may be considered a useful tool for HPV molecular diagnosis.

Stability of human papillomavirus (HPV) in cervical ThinPrep specimens previously lysed with glacial acetic acid: effect on cobas 4800 HPV test performance.

BACKGROUND: The authors previously demonstrated that lysing samples with glacial acetic acid (GAA) before human papillomavirus (HPV) testing does not adversely affect HPV detection with the cobas 4800 HPV Test. However, the long-term impact of GAA on HPV DNA was not explored in that study, and inherent cell loss with the lysing protocol used also was observed. The current study considered the long-term effects of GAA treatment of cervical ThinPrep samples on HPV detection with the cobas 4800 HPV Test. They also modified the manufacturer's lysing procedure for ThinPrep specimens to help prevent cell loss. METHODS: Seventy-eight ThinPrep samples, including previously lysed, archived specimens, were split; then, 1 part was treated with GAA according to the manufacturer's protocol or using a modified protocol, and the other part was left untreated. All samples were tested for HPV using the cobas 4800 HPV Test. The HPV-positive/HPV-negative status of tested samples was used to determine the level of agreement between treated and untreated fractions. Performance of the modified lysing procedure was assessed relative to the manufacturer's protocol. RESULTS: Positive HPV status produced 97% agreement between the GAA-treated/untreated fractions. Concordance between the GAA-treated/untreated fractions did not differ between the 2 lysing methods; however, the average percentage increases in ß-globin and HPV cycle threshold values after lysing were less discernible for the modified lysing method. CONCLUSIONS: GAA treatment of cervical ThinPrep specimens does not have long-term adverse effects on HPV detection with the cobas 4800 HPV Test. The manufacturer's GAA lysing procedure for ThinPrep specimens can be reliably modified for specimens that may subsequently undergo HPV testing.

Detección molecular del papilomavirus humano de alto riesgo en el seguimiento de mujeres tratadas por lesión escamosa intraepitelial / Molecular detection of high risk human papillomavirus in following women treated for squamous intraepithelial lesion

El cáncer de cuello uterino (CCU) es la primera causa de muerte por cáncer en mujeres en países en vías de desarrollo. La infección persistente por el virus papiloma humano (VPH) es un factor necesario en lesiones preneoplásicas y CCU. La citología cervicovaginal es el método mayormente utilizado para detectar el CCU y su uso combinado con la de detección de ADN viral seis meses post-tratamiento aumenta la efectividad para identificar mujeres tratadas con riesgo de lesión residual/recidiva. El objetivo fue describir la frecuencia de VPH de alto riesgo (AR) en mujeres tratadas por lesión escamosa intraepitelial (SIL) que acudieron al Servicio de Patología Cervical del Hospital San Pablo de enero-diciembre/2014. Se realizó un estudio descriptivo de corte transverso, que incluyó 80 pacientes que acudieron al servicio para control post-tratamiento por SIL. Se utilizó Cobas 4800 HPV Test (Roche) para la detección individual de VPH-16 y 18, y un pool de 10 VPH-AR (31,33,35,39,45,51,52,56,58,59) y dos de “probable” alto riesgo (66,68). Se encontró infección viral en el 7,5% (6/80) de las pacientes tratadas; identificándose VPH-16 en 3/6 de los casos positivos. En Paraguay existe alta incidencia de lesiones pre-neoplásicas y CCU siendo un problema de salud pública. Los datos observados sugieren que la utilización de este sistema para la detección viral puede llevar a optimizar el seguimiento post-tratamiento y la identificación de VPH-16 y 18 podría contribuir a la selección de pacientes en mayor riesgo de desarrollar una lesión cervical que deben someterse a una vigilancia frecuente y meticulosa.

Cervical cancer is the leading cause of cancer death in women in developing country.Persistent infection with human papillomavirus (HPV) is a necessary factor in premalignantlesions and cervical cancer. The Pap smear is the method most commonly used to detect thecervical cancer and its combined with the detection of viral DNA six months post-treatment increases effectiveness to identify women treated in risk with residual/recurrent lesion. Theaim was to describe the frequency of high-risk HPV (HR) in treated women for squamousintraepithelial lesion (SIL) who attended the Cervical Pathology Service of Hospital San Pablofrom January to December/2014. It is cross sectional, descriptive study which included 80patients who attended the service for post-treatment control. The 4800 Cobas HPV Test(Roche) was used, which detects HPV-16 and 18, and a pool of 10 HR-HPV(31,33,35,39,45,51,52,56,58,59) and two "probable" high risk (66,68). Viral infection wasfound in 7.5% (6/80) of patients treated; identifying HPV-16 in 3/6 of positive cases. InParaguay there is a high incidence of pre-neoplastic lesions and cervical cáncer remains apublic health problem. The observed data suggest that the use of this system for viraldetection can lead to optimize the post-treatment monitoring and the identification of HPV-16and 18 could contribute to the selection of patients at increased risk of developing cervicalinjury should undergo to frequent and careful monitoring.

Detección molecular del virus papiloma humano de alto riesgo oncogénico en muestras cervicales. laboratorio central de salud pública. primeros resultados / Molecular detection of high oncogenic risk human papiloma virus in cervical samples. central laboratory of public health. first results

El cáncer de cuello uterino es la primera causa de muerte por cáncer en mujeres en países en vías de desarrollo, con una tasa de incidencia de 34,2 por 100.000 mujeres y de mortalidad de 15,7 por 100.000 mujeres en Paraguay. La sensibilidad de la citología está entre 30-60%, mientras que la de la detección molecular del Virus Papiloma Humano (VPH) en muestras cervicales, es mayor al 90% para detectar neoplasia intraepitelial cervical de grado 2 (CIN II) o más. El objetivo de este trabajo fue describir la frecuencia de detección de VPH de alto riesgo (AR) y su distribución por edad en mujeres que concurrieron al Hospital San Pablo, de mayo a agosto de 2.013. Se estudiaron 170 muestras cervicales de pacientes que accedieron a participar firmando un consentimiento informado. Se utilizó el sistema Cobas 4800 HPV Test(Roche) que detecta los VPH-AR 16 y 18, y un pool de 10 VPH-AR (31,33,35,39,45,51,52,56,58,59) y dos de "probable" alto riesgo (66,68). La frecuencia de infección por VPH-AR fue del 16%, la infección decrecía con la edad y el mayor número de casos apareció en mujeres menores de 30 años. El VPH-16 fue encontrado en todos los grupos de edades. Este es el primer reporte de la detección de ADN de VPH-AR en el LCSP, y se muestra que la prevención y control del cáncer cérvico-uterino es una prioridad de salud pública en el país por la gran carga de la enfermedad evidenciada por su alta incidencia y mortalidad.

Cervical cancer is the leading cause of cancer death in women in developing countries,with an incidence rate of 34.2 per 100,000 women and 15.7 deaths per 100,000 womenin Paraguay. The sensitivity of cytology is 30-60% while molecular detection of HumanPapilloma Virus (HPV) in cervical samples is over 90% for cervical intraepithelial neoplasiagrade 2 (CIN II) or more. The aim of this study was to describe the frequency ofdetection of high-risk HPV (HR) and their distribution by age in women who attended theHospital San Pablo, from May to August 2013. One hundred seventy cervical samplesfrom patients who agreed to participate by signing an informed consent. The 4800 Cobassystem HPV Test (Roche), which detects the HR-HPV genotypes 16 and 18, a pool of 10 HR-HPV (31,33,35,39,45,51,52,56,58,59) and two "probable" high risk (66.68). Thefrequency of HR-HPV infection was 16%, the infection decreased with age and the highestnumber of cases occurred in women under 30 years. HPV-16 was found in all age groups.This is the first report about detection of DNA HR-HPV in the LCSP and it showed thatprevention and control of cervical cancer is a public health priority in the country by thelarge burden of the disease as evidenced by its high incidence and mortality.

Prevalence of high-risk human papillomavirus by cobas 4800 HPV test in urban Peru

Background: Molecular tests allow the detection of high-risk human papillomavirus in cervical samples, playing an important role in the prevention of cervical cancer. Objectives: We performed a study to determine the prevalence of HPV 16, HPV 18 and other high-risk human papillomavirus (pool 12 genotypes) in Peruvian females from diverse urban areas using the cobas 4800 HPV test. Methods: Routine cervical samples collected in our laboratory were analyzed by cobas 4800 HPV test. Results: A total of 2247 samples from female patients aged 17–79 years were tested. high-risk human papillomavirus was positive in 775 (34.49%) samples. Of these, 641 (82.71%) were single infections and 134 (17.29%) were multiple infections. The positivity rates for HPV 16, HPV 18, and other high-risk human papillomavirus were 10.77%, 2.0%, and 28.08%, respectively. In multiple high-risk human papillomavirus infections, the concomitance of HPV 16 and other high-risk human papillomavirus was more prevalent (13.42%). Conclusion: Our study showed high prevalence of high-risk human papillomavirus in urban Peru, mainly among young women. In both single and multiple infections other high-risk human papillomavirus were more prevalent than HPV 16 and HPV 18, which might influence vaccine impact in our country. Furthermore, the cobas 4800 HPV test may be considered a useful tool for HPV molecular diagnosis. .