A Comparison between Real-Time Polymerase Chain Reaction and Hybrid Capture 2 for Human Papillomavirus DNA Quantitation

• Published in: Cancer epidemiology, biomarkers & prevention biomarkers & prevention, 2003-06, Vol.12 (6), p.477-484
• Main Authors: GRAVITT, Patti E, BURK, Robert D, LORINCZ, Attila, HERRERO, Rolando, HILDESHEIM, Allan, SHERMAN, Mark E, CONCEPCION BRATTI, Maria, RODRIGUEZ, Ana Cecilia, HELZLSOUER, Kathy J, SCHIFFMAN, Mark
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Carcinoma, Squamous Cell - genetics; Carcinoma, Squamous Cell - virology; Case-Control Studies; Cervical Intraepithelial Neoplasia - genetics; Cervical Intraepithelial Neoplasia - virology; Cohort Studies; Computer Systems; Costa Rica - epidemiology; Cross-Sectional Studies; DNA, Viral - analysis; DNA, Viral - genetics; DNA, Viral - isolation & purification; Female; Female genital diseases; Genotype; Gynecology. Andrology. Obstetrics; Humans; Medical sciences; Nucleic Acid Hybridization; Oncogene Proteins, Viral - analysis; Oncogene Proteins, Viral - genetics; Oncogene Proteins, Viral - isolation & purification; Papillomaviridae - genetics; Papillomaviridae - isolation & purification; Polymerase Chain Reaction - methods; Predictive Value of Tests; Prevalence; Severity of Illness Index; Statistics as Topic; Tumor Virus Infections - genetics; Tumor Virus Infections - virology; Tumors; Uterine Cervical Neoplasms - genetics; Uterine Cervical Neoplasms - virology; Viral Load; Women's Health
• ISSN: 1055-9965; 1538-7755

Studies investigating human papillomavirus (HPV) viral load as a risk factor in the development of squamous intraepithelial lesions (SILs) and cancer have often yielded conflicting results. These studies used a variety of HPV viral quantitation assays [including the commercially available hybrid capture 2 (HC 2) assay], which differ in their ability to account for differences in cervical cell collection, linear dynamic range of viral load quantitation, and determination of type-specific versus cumulative viral load measures. HPV-16 and HPV-18 viral quantitation using real-time PCR assays were performed to determine whether type-specific viral load measurements that adjust for specimen cellularity result in a different association between viral load and prevalent SIL and cancer, compared with HC 2 quantitation (which does not adjust for cellularity or multiple infections). In general, HPV-16 viral load as measured by real-time PCR increased linearly with increasing grade of SIL while HPV-18 measured using similar techniques increased through low-grade SIL (LSIL), with HPV-18 viral load among high-grade SIL and cancers near the level of cytologically normal women. HC 2 viral load, using the clinical 1.0 pg/ml cut point, differentiated cytologically normal women from women with any level of cytological abnormality (normal versus ≥LSIL) but did not change as lesion severity increased. There was no evidence for plateau of HC 2 at high copy numbers, nor was significant variability in total specimen cellularity observed. However, cumulative viral load measurements by HC 2, in the presence of multiple coinfections, overestimated type-specific viral load. Multiple infections were more common among women with no (32%) or LSIL (51%) [ versus 23% in high-grade SIL/cancer], partially explaining the lack of a dose response using a cumulative HC2 viral load measure. The nonrandom distribution of multiple infections by case-control status and the apparent differential effect of viral load by genotype warrant caution when using HC 2 measurements to infer viral load associations with SIL and cancer.