High agreement of Seegene Anyplex™ HPV 28 and Allplex™ HPV 28 for high-risk vaccine-type HPV detection in the KEN SHE Study.

BACKGROUND

The KEN SHE Study replaced Anyplex™ II HPV28 Detection multiplex real-time PCR kits (Anyplex) with Allplex™ HPV28 kits (Allplex) mid-study. Assay comparison is necessary for ongoing research.

OBJECTIVES

To estimate agreement between Allplex and Anyplex assays for vaccine type HPV DNA detection within the KEN SHE Study.

STUDY

DESIGN

The KEN SHE Study is a randomized clinical trial of single-dose HPV vaccination. HPV DNA measured in cervical swab specimens collected every six months is used to ascertain the study endpoint of persistent positive HPV. Two hundred specimens previously tested using Anyplex were randomly selected for re-assay using Allplex. Prespecified selection criteria ensured high positivity for HPV 16/18/31/33/45/52/58 (70% positive per Anyplex). Cohen's Kappa coefficients were used to assess inter-assay agreement.

RESULTS

Agreement between Anyplex and Allplex was κ= 0.93 (95%CI: 0.88-0.99) for HPV 16, κ= 0.96 (0.91-1.00) for HPV 18, κ= 0.97 (95% CI: 0.90-1.00) for HPV 31, κ= 0.93 (0.80-1.00) for HPV 33, κ= 0.65 (0.37, 0.94) for HPV 45, κ= 0.88 (0.79-0.97) for HPV 52, and κ= 0.92 (0.84-1.00) for HPV 58. Inter-assay agreement for combined vaccine types HPV 16/18, HPV 31/33/45/52/58, and HPV 16/18/31/33/45/52/58 was also high. Most discordant results either had high cycle threshold (ct) values on Allplex (ct>35) or a single semiquantitative '+ ' on Anyplex.

CONCLUSIONS

The Allplex assay has excellent agreement with Anyplex for individual HPV types 16, 18, 31, 33, 52, and 58, as well as combined HPV 16/18, HPV 16/18/31/33/45/52/58 and HPV 31/33/45/52/58. These results support continued analysis with the Allplex assay.