A comparison between real-time polymerase chain reaction and hybrid capture 2 for human papillomavirus DNA quantitation

Patti E. Gravitt, Robert D. Burk, Attila Lorincz, Rolando Herrero, Allan Hildesheim, Mark E. Sherman, Maria Concepcion Bratti, Ana Cecilia Rodriguez, Kathy J. Helzlsouer, Mark Schiffman

Research output: Contribution to journalArticlepeer-review

110 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)477-484
Number of pages8
JournalCancer Epidemiology Biomarkers and Prevention
Issue number6
StatePublished - Jun 1 2003
Externally publishedYes

ASJC Scopus subject areas

  • Epidemiology
  • Oncology


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