Comparison of Analytical and Clinical Performance of HPV 9G DNA Chip, PANArray HPV Genotyping Chip, and Hybrid-Capture II Assay in Cervicovaginal Swabs

BACKGROUND: Human papillomavirus (HPV) infection can be detected by using several molecular methods, including Hybrid-Capture II (HC2) assay and variable HPV DNA chip tests, although each method has different sensitivities and specificities. METHODS: We performed HPV 9G DNA Chip (9G) and PANArray HPV Genotyping Chip (PANArray) tests on 118 cervicovaginal swabs and compared the results with HC2, cytology, histology, and direct sequencing results. RESULTS: The overall and high-risk HPV (HR-HPV) positivity rates were 62.7% and 44.9% using 9G, and 61.0% and 30.5% using PANArray, respectively. The positivity rates for HR-HPV with these two chips were significantly lower than 55.1% when HC2 was used. The sensitivity of overall HPV positivity in detecting histologically confirmed low-grade cervical squamous intraepithelial lesions or higher was 88.7% for all three tests. The specificity was 58.5% for 9G and 61.5% for PANArray, which was significantly lower than the 72.3% for HC2. With the HR-HPV+ genotype threshold, the sensitivity decreased to 75.5% for 9G and 52.8% for PANArray, which was significantly lower than the 88.7% for HC2. Comparison of the two chips showed concordant results in 55.1% of the samples, compatible results in 16.9%, and discordant results in 28.0%, exhibiting poor agreement in detecting  certain HPV genotypes. Compared with direct sequencing, 9G yielded no discordant results, whereas PANArray yielded 31 discordant results (26.7%). CONCLUSIONS: Compared with HC2, the HPV genotyping tests showed lower sensitivity in histologic correlation. When the two chips were compared, the 9G was more sensitive and accurate for detecting HR-HPV than the PANArray.

Detección del virus del papiloma humano de alto riesgo por captura híbrida II® según hallazgos citológicos en mujeres tratadas por lesiones escamosas intraepiteliales de cuello uterino, período 2006/2010 / Detection of high risk human papillomavirus by hybrid capture II(r) according cytological findings in women treated for squamous intraepithelial lesions of the cervix, period 2006/2010

OBJETIVO: Determinar la frecuencia del virus de papiloma humano de alto riesgo oncogénico (HR-HPV) por captura híbrida II (r) (CH II(r)) según hallazgos citológicos en mujeres tratadas por lesiones escamosas intraepiteliales (SIL) de cuello uterino. MATERIAL Y MÉTODO: Estudio descriptivo de corte transverso de una serie de casos, en donde se incluyeron 122 mujeres tratadas, 79 (65%) por SIL de bajo grado (LSIL) y 43 (35%) por SIL de alto grado (HSIL) que concurrieron al Laboratorio de HPV del Instituto de Investigaciones en Ciencias de la Salud, Universidad Nacional de Asunción, para realizarse un control post-tratamiento, periodo 2006/2010. RESULTADOS: Se observó un total del 28% (34/122) de mujeres tratadas por SIL positivas para HR-HPV, detectándose infección viral en un 20% de las mujeres con ausencia de SIL (NSIL) (22/108), 83% de las mujeres con LSIL (10/12) y 100% de las mujeres con HSIL (2/2). De las 34 mujeres positivas para HR-HPV, 10 mujeres (29%) presentaron valores altos (100 pg/mL o más) de carga viral relativa, detectándose un aumento de casos positivos con la severidad de la lesión (28% NSIL, 30% LSIL, 50% HSIL). CONCLUSION: La detección de HR-HPV por CH II(r), así como los valores de carga viral relativa altos, en especial en mujeres con NSIL podrían ayudar a identificar mujeres tratadas con riesgo a desarrollar recidivas, contribuyendo así a fortalecer el programa de prevención de cáncer de cuello uterino. .

OBJECTIVE: To determinate the frequency of high risk human papillomavirus (HR-HPV) by hybrid capture II (r) (CH II(r)), according cytology results in women treated for squamous intraepithelial lesions of the cervix (SIL). MATERIAL AND METHODS: A descriptive cross-sectional study of a series of cases that included 122 women treated, 79 (75%) for low grade SIL (LSIL) and 43 (35%) for high grade SIL (HSIL) attending at the HPV Laboratory at the Health Sciences Research Institute (IICS), National University of Asunción (UNA), for post-treatment control during period 2006/2010. RESULTS: A total of 28% (34/122) of women treated for SIL were positive for HR-HPV, detecting viral infection in 20% of women with no SIL (NSIL) (22/108), in 83% of women with LSIL (10/12) and in 100% of women with HSIL (2/2). Of 34 women positive for HR-HPV, 10 women (29%) had high values (100 pg / mL or more) of relative viral load, detecting an increase of positive cases with severity of the lesion (28% NSIL, 30% LSIL, 50% HSIL). CONCLUSION: HR-HPV detection by CH II(r) and high relative viral load values especially in women with NSIL could help to identify treated women at risk of developing recurrence, thereby contributing to strengthening the cervical cancer prevention program. .

Frecuencia y carga viral relativa del virus del papiloma humano de alto riesgo según el diagnóstico citológico en mujeres paraguayas por captura híbrida II / Frequency and relative viral load of high risk human papilloma virus according to the cytological diagnosis in paraguayan women by hybrid capture II

El cáncer de cuello uterino es el tumor maligno más frecuente en mujeres de Latinoamérica y su agente causal es el virus del papiloma humano (HPV). Recientemente en Paraguay incorporamos el método de captura híbrida II (CH II), el cual detecta 13tipos de HPV de alto riesgo oncogénico (HR-HPV) y proporciona valores relativos de carga viral. El objetivo del estudio fue determinar la carga viral relativa de HR-HPV por CH IIsegún el diagnóstico citológico. Fueron incluidas 566 mujeres (33 + 10 años) atendidas en el Instituto de Investigaciones en Ciencias de la Salud 2006/2009. Fue detectado HRHPVen 43% de las mujeres (241/566), observándose una alta frecuencia del 23% en mujeres con ausencia de lesión intraepitelial (NSIL). Según el diagnóstico citológico, se evidenció una diferencia altamente significativa entre los valores de carga viral relativa(p<0,0001; Kruskal Wallis), observándose un aumento de carga viral relativa de mujeres con NSIL a mujeres con SIL (68 pg/ml en ausencia de SIL; 710 pg/ml para SIL de bajo grado-LSIL y 474 pg/ml para SIL de alto grado-HSIL). No se observó cambio significativo en la carga viral relativa entre LSIL y HSIL (p=0,60; prueba t de Student). Los resultados sugieren que los valores de carga viral relativa proporcionados por CH II pueden ser considerados como un indicador importante en el manejo de mujeres con sospecha de SIL.

Cervical cancer is the most frequent malignant tumor in women of Latin America being human papillomavirus (HPV) the main causative agent. Recently in Paraguay, we incorporated the method of hybrid capture II (CH II) which detects 13 types of HPV ofhigh oncogenic risk (HR-HPV) and provides relative values of viral load. The aim of this study was to determine the frequency and relative viral load of HR-HPV by CH II according to the cytological diagnosis. There were 566 women (33 + 10 years) included in the study that attended the Research Institute in Health Sciences during the period 2006/2009. HR-HPV was detected in 43% of the women (241/566), being observed a high frequency of 23% in women negative to squamous intraepithelial lesions (NSIL). According to the cytological diagnosis, there was a highly significant difference between the values of relative viral load (p <0.0001; Kruskal Wallis), with an increase of relative viral load of women with NSIL to women with SIL (mean values: 68 pg/ml in NSIL; 710 pg/ml in low grade SIL-LSIL and 474 pg/ml in high grade SIL-HSIL). Significant change was not observed in the relative viral load between LSIL and HSIL (p=0.60; Student’s t test).The results suggest that the values of relative viral load provided by CH II could be considered an important indicator for managing women with suspicion of SIL.

Detection of high risk human papillomavirus cervical infections by the hybrid capture in Asunción, Paraguay

Cervical cancer is the most frequent malignant tumour of women in Latin America being human papillomavirus (HPV) the main cause. The aim of this study was to increase the knowledge about the cervical infections with oncogenic HPV types (HR-HPV) in Asuncion, Paraguay. Two hundred and seventy-two cervical samples were analyzed using hybrid capture II assay (HCA II) for HR-HPV. The frequency of HR-HPV in the study group was 44 percent. HR-HPV was detected in 25 percent of the women negative for squamous intraepithelial lesions (NSIL), 72 percent with atypical squamous cells of undetermined significance (ASCUS), 68 percent with low SIL and 78 percent with high SIL. A moderate concordance was observed between HCA II assay and cytology (kappa: 0.43 IC95 percent 0.3 - 0.5). It was detected a high frequency of HR-HPV in women from 11 to 30 years old and in those over 60 years old. The data obtained in this study showed a high frequency of HR-HPV in woman with NSIL and ASCUS, which corroborate that the use of cytology together with HCA II assay for HR-HPV could improve remarkably the efficiency of screening programs of cervical cancer in Paraguay. Furthermore, these findings point out the need for the periodical follow-up of HR-HPV infections in older women.

A Comparision of Surepath(TM) Liquid-Based Smear with a Conventional Smear for Cervicovaginal Cytology-with Reference to a Histological Diagnosis / 대한세포병리학회지

This study was performed to compare Surepath(TM) liquid-based smear and a conventional cervicovaginal smear with reference to a histological diagnosis. A hybrid capture test (HCII) was also performed and analyzed. We collected matched cases for cervicovaginal cytology- histology: 207 cases for conventional cytology (CC) and 199 cases for liquid-based cytology (LBC). HCII was performed in 254 patients. When a cytological diagnosis of ASCUS or above (ASCUS+) is classified as positive and a histological diagnosis of LSIL+ is classified as positive, the sensitivity and specificity for LBC was 91.7% and 75.9%, respectively and the sensitivity and specificity for CC was 62.6% and 96.1%, respectively. When a cytological and histological diagnosis of LSIL+ is classified as positive, the sensitivity and specificity for LBC was 77.5 and 96.6%, respectively and the sensitivity and specificity for CC was 49.7% and 100%, respectively. When a histological diagnosis of LSIL+ is classified as positive, the sensitivity and specificity for HCII was 78.9% and 78.1%, respectively. The concordance ratio between the cytological and histological diagnosis was 80.4% (kappa=76.0) for LBC and 56.5% (kappa=55.1) for CC. LBC is more sensitive and less specific then CC, as a cytological cutoff level of ASCUS, but more sensitive and equally specific, as a cytological cutoff level LSIL or HSIL. LBC is more reliable with a high concordance ratio between the cytological and histological diagnosis.

Evaluation of HR-HPV testing in diagnosis of cervical intraepithelial neoplasia and cervical cancer / 临床检验杂志

Objective To evaluate the role of high-risk human papillomavirus (HR-HPV) testing in diagnosis of high-grade cervical intraepithelial neoplasia (CIN2/3+) and cervical cancer.Methods Total 3426 women with known cytologic abnormality tests were subjected to detect high-risk HPV DNA by hybrid capture II and to have established diagnoses by cervical coloposcopy-directed biopsy test.Results Among 3426 women,45 were cervical cancer,670 were CIN 2/3,and 2711 were CIN 1 or other benign lesions based on the cervical histologic results. Compared with the women with CIN 1 or other benign lesions,the women with CIN 2/3+ had higher positive rates of HR-HPV. Higher load of HR-HPV was more common among women with cervical lesions than those without. Fifty-seven women who were transiently HR-HPV positive did not develop CIN2/3+ during follow-up.Conclusions Our data suggest that a high viral load could be used as a short-term marker of CIN 2/3+,and women repeatedly tested positive for HR-HPV are at risk of developing CIN2/3+. Detecting HR-HPV and follow up the positive women would be useful in diagnosis of CIN2/3+.

The significance of Human Papillomavirus viral load by using hybrid capture II assay in diagnosis of CIN II or above in women with PAP smear showing only ASCUS/LSIL / 대한산부인과학회지

OBJECTIVE: To determine the clinical significance of high risk Human papillomavirus (HPV) DNA viral load measured by hybrid capture (HC) II assay in diagnosis of CIN II or above in women with cervical smear showing only Atypical squamous cells of undetermined significance (ASCUS)/Low-grade squamous intraepithelial lesion (LSIL). METHODS: This study was performed from September 2003 to September 2004 and included 111 patients with ASCUS/LSIL on PAP smear. All of them underwent high risk HPV DNA tests by HC II assay and colposcopy directed cervical biopsies. The histologic results were classified as low grade lesion (normal, CIN I) and high grade lesion (CIN II, III, and invasive cancer). The viral load was transformed into their logarithm (Log10RLU/PC ratio), and categorized as following four groups; negative HPV DNA (Log10RLU/PC or =2.0). We evaluated the relationships of the detection rate and viral load of high risk HPV DNA with histologic severity of cervical lesions. RESULTS: The rate of CIN II or above in 111 patients with ASCUS/LSIL was 25.2%. The detection rate of high risk HPV DNA using HC II assay was 96.4% in high grade lesion, and 66.3% in low grade lesion of histologic test. There was strong correlation between CIN II or above and positivity for HPV DNA when this group was compared with women with only CIN I or normal cervix (OR:13.7, 95% CI:1.7-106.4). The higher the viral load of HPV DNA infection observed, the higher the probability of being associated with stage of CIN (correlation coefficient:0.279). The frequency of high viral load observed in high grade lesion (89.3%) was significantly higher than low grade lesion (33.7%)(OR:16.3, 95% CI:4.5-58.9). When the cutoff of positivity was changed from 1.0RLU/PC ratio to 100.0RLU/PC ratio (Log10 RLU/PC ratio=2.0), the sensitivity for the prediction of high grade lesion was slightly decreased (96.4->89.3%), but pronounced gain in specificity (33.7->66.3%), and reduction in false positive rate (66.3->33.7%). CONCLUSION: The viral load of high risk HPV DNA measured by HC II assay was significantly associated with the severity of histology, and useful in diagnosis of CIN II or above in women referred because of ASCUS/LSIL on PAP smear.