Analytical characterization of the APTIMA HPV Assay

J Clin Virol. 2009 Jul:45 Suppl 1:S39-47. doi: 10.1016/S1386-6532(09)70007-1.

Abstract

Background: Human papillomavirus (HPV) testing has improved the sensitivity for the detection of cervical pre-cancer and cancer as compared to Pap testing. Several HPV tests are commercially available and most target the DNA from 13 or 14 high-risk HPV types. The APTIMA HPV Assay however, detects HPV E6/E7 mRNA from 14 high-risk types of HPV: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68.

Objective: To determine the analytical performance characteristics of the APTIMA HPV Assay.

Study design: Analytical sensitivity, analytical specificity, reproducibility, and the effect of potentially interfering substances was determined for the APTIMA HPV Assay on both the DTS (semi-automated) and TIGRIS DTS (fully automated) systems.

Results: The 95% detection limit for both systems was between 17 and 488 copies/reaction, depending on the HPV type. The assay did not cross-react with normal flora and opportunistic organisms that may be found in cervical samples, or low-risk HPV types. Spermicides, anti-fungal and anti-itch medications, whole blood, glacial acetic acid, and most lubricants did not interfere with assay performance. Those lubricants containing polyquaternium 15 did interfere with assay performance. Inter-instrument, inter-operator, inter-lot, and inter-run signal variability were <10% for >99% of the data. Intra-run variability was <15%, except for those samples with concentrations at or below the 95% detection limit of the assay.

Conclusions: Based upon the analytical sensitivity, analytical specificity, and low variability, the APTIMA HPV Assay showed excellent performance and robustness.

Publication types

  • Evaluation Study

MeSH terms

  • Automation
  • Cervix Uteri / virology
  • Female
  • Gene Expression Profiling / methods*
  • Humans
  • Oncogene Proteins, Viral / biosynthesis*
  • Oncogene Proteins, Viral / genetics
  • Papillomaviridae / pathogenicity*
  • Papillomavirus Infections / virology*
  • RNA, Messenger / biosynthesis*
  • RNA, Messenger / genetics
  • RNA, Viral / biosynthesis*
  • RNA, Viral / genetics
  • Reagent Kits, Diagnostic*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Uterine Cervical Neoplasms / diagnosis*

Substances

  • Oncogene Proteins, Viral
  • RNA, Messenger
  • RNA, Viral
  • Reagent Kits, Diagnostic