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Dual-Staining Outperforms Cytology for Follow-up of HPV-positive Women in Cervical Cancer Screening Programs

, by DCEG Staff

Dual stain testing on Pap test samples identifies the presence of two proteins: p16 (brown) and Ki-67 (red).

In a population of women undergoing routine cervical cancer screening, dual staining for cellular proteins p16/Ki-67 outperformed cytology (evaluation of cells under the microscope) in detecting precancer among human papillomavirus (HPV)-positive women. At the same time, dual staining referred fewer women to colposcopy, a secondary screening involving visual inspection of the cervix with magnification. These findings, published May 13, 2019, in the journal JAMA Internal Medicine, build upon the prior proof-of-principle testing of dual staining as a biomarker of cervical precancers.

Nicolas Wentzensen, M.D., Ph.D., M.S., and colleagues carried out this research in a cohort of over 3,000 HPV-positive women ages 25-65 undergoing HPV and Pap cytology testing in the Kaiser Permanente Northern California (KPNC) system. Because HPV infection is ubiquitous in the population, screening programs that utilize primary HPV screening need effective ways to triage HPV-positive women, most of whom experience transient infections that will not progress to cervical precancer. Though Pap cytology successfully identifies women with precancer, a lot of women without precancer are sent for additional follow-up, which is costly, time-consuming, and can result in unnecessary cervical biopsies. In the latest analysis, dual staining was shown to be as effective as Pap cytology at identifying cervical precancer, while at the same time, dramatically reducing the number of women sent to follow-up. The authors conclude that “dual staining can safely replace Pap cytology as a triage strategy for primary HPV screening.”

The assay is currently under review by the U.S. Food and Drug Administration.


Wentzensen, N, Clarke, M, et al. Clinical evaluation of HPV screening with p16/Ki-67 dual stain triage in a large organized cervical cancer screening program. JAMA Internal Medicine. May 13, 2019. DOI:10.1001/jamainternmed.2019.0306 [Epub ahead of print]

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