Viral Load of High-Risk Human Papillomaviruses as Reliable Clinical Predictor for the Presence of Cervical Lesions

Abstract

Background: Infections with high-risk human papillomaviruses (Hr-HPV) can cause malignant transformation of the human cervical epithelium. HPV DNA tests generally are very sensitive to detect cervical neoplastic lesions but also identify transient HPV infections. As a consequence, the specificity and positive predictive value are low.Methods: We analyzed viral load of Hr-and possibly Hr-HPV types more than seven orders of magnitude (on a log10 scale) in 999 consecutive BD-SurePath liquid-based cervical cytology samples from routine cervical screening enriched with atypical squamous cells of undetermined significance (n ¼ 100), low-grade squamous intraepithelial lesions (LSIL; n ¼ 100), and high-grade squamous intraepithelial lesions (HSIL; n ¼ 97) using type-specific multiplex quantitative real-time PCR and the BSGP5þ/6þ-PCR/MPG assay. In the 36-month follow-up, 79 histologically verified CIN2þ and 797 double-negative cytology cases were identified.Results: Viral loads in LSIL and HSIL were significantly increased compared with no intraepithelial lesion or malignancy in both the quantitative PCR (qPCR) and BSGP5þ/6þ-PCR/MPG assay (P < 0.0001). The mean viral loads in LSIL and HSIL were not significantly different. Using a newly determined high viral load cut off for 14 Hr-HPV types, the sensitivity for prevalent CIN3þ remained at 100% for both assays compared with the minimal detection threshold. The specificity (corresponding to double-negative cytology at subsequent screening episodes) increased substantially (qPCR, from 91.1% to 95.7%; BSGP5þ/6þ-PCR/MPG, from 79.8% to 96.2%).Conclusions: Compared with DNA positivity alone, high Hr-HPV viral loads could reduce the amount of false positive results detected by the BSGP5þ/6þ-PCR/MPG and qPCR by 81.4% and 52.1%, respectively.Impact: Quantitative type-specific HPV DNA assays show high flexibility in defining thresholds that allow optimizing clinical accuracy for cervical cancer precursors. Cancer Epidemiol Biomarkers Prev; 22(3); 406-14. Ó2013 AACR.

Multiple Human Papillomavirus Infections with High Viral Loads Are Associated with Cervical Lesions but Do Not Differentiate Grades of Cervical Abnormalities

“…The current study is part of the VALGENT project, which provides a comprehensive design to validate and compare general HPV tests (identifying Hr-HPV infections) and HPV genotyping assays (identifying some or all high-risk HPV types separately) (12)(13)(14). Starting 1 October 2006, 999 consecutive cytology samples were collected during routine gynecological health checks from women in Flanders, Belgium.…”

“…All samples were collected with a Cervex-Brush Combi and prepared using BD SurePath technology for liquid-based cytology (15,16). The study samples were supplemented by smears from patients with atypical squamous cells of undetermined significance (ASCUS) (n ϭ 100), LSIL (n ϭ 100), and HSIL (n ϭ 97), as described recently (13,14). Study-specific patient identification codes were assigned and transmitted in such a manner that patient confidentiality was preserved.…”

“…Thin-layer slide preparations were made with the fully robotic AutoCyte PREP System7 (Tripath Imaging, Inc.) and were prepared as described previously (17). HPV-targeted cytology was read on all cytology samples with prior knowledge of quantitative PCR (qPCR) HPV DNA data (14). The cytological results were classified according to the 2001 Bethesda System (18).…”

“…The HPV genotyping and ␤-globin assessment by the BSGP5 ϩ /6 ϩ -PCR/MPG assay were performed as described previously (10,14,21,22). HPV amplification was carried out using the Multiplex PCR kit (Qiagen, Hilden, Germany).…”

Cancer and the microbiome: potential applications as new tumor biomarker

“…One way to achieve this goal would be to routinely search for specific pro-oncogenic pathogens in all patients diagnosed with certain cancer types within a multidisciplinary, multicenter and multinational consortium program. That approach has been already used by the VALGENT study group in the case of the research that correlates the HPV infection and the presence of cervical lesions [132,133]. That enormous effort may definitely contribute to a better knowledge on how different microbiome alterations can facilitate or prevent cancer development.…”