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 journalArticle

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Abstract

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
Volume12
Issue number6
StatePublished - Jun 1 2003

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Viral Load
Real-Time Polymerase Chain Reaction
DNA
Human papillomavirus 18
Human papillomavirus 16
Neoplasms
Infection Control
Infection
Squamous Intraepithelial Lesions of the Cervix
Coinfection
Genotype

ASJC Scopus subject areas

  • Epidemiology
  • Oncology

Cite this

Gravitt, P. E., Burk, R. D., Lorincz, A., Herrero, R., Hildesheim, A., Sherman, M. E., ... Schiffman, M. (2003). A comparison between real-time polymerase chain reaction and hybrid capture 2 for human papillomavirus DNA quantitation. Cancer Epidemiology Biomarkers and Prevention, 12(6), 477-484.

A comparison between real-time polymerase chain reaction and hybrid capture 2 for human papillomavirus DNA quantitation. / Gravitt, Patti E.; Burk, Robert D.; Lorincz, Attila; Herrero, Rolando; Hildesheim, Allan; Sherman, Mark E.; Bratti, Maria Concepcion; Rodriguez, Ana Cecilia; Helzlsouer, Kathy J.; Schiffman, Mark.

In: Cancer Epidemiology Biomarkers and Prevention, Vol. 12, No. 6, 01.06.2003, p. 477-484.

Research output: Contribution to journalArticle

Gravitt, PE, Burk, RD, Lorincz, A, Herrero, R, Hildesheim, A, Sherman, ME, Bratti, MC, Rodriguez, AC, Helzlsouer, KJ & Schiffman, M 2003, 'A comparison between real-time polymerase chain reaction and hybrid capture 2 for human papillomavirus DNA quantitation', Cancer Epidemiology Biomarkers and Prevention, vol. 12, no. 6, pp. 477-484.
Gravitt, Patti E. ; Burk, Robert D. ; Lorincz, Attila ; Herrero, Rolando ; Hildesheim, Allan ; Sherman, Mark E. ; Bratti, Maria Concepcion ; Rodriguez, Ana Cecilia ; Helzlsouer, Kathy J. ; Schiffman, Mark. / A comparison between real-time polymerase chain reaction and hybrid capture 2 for human papillomavirus DNA quantitation. In: Cancer Epidemiology Biomarkers and Prevention. 2003 ; Vol. 12, No. 6. pp. 477-484.
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