Comparison of three different PCR methods for quantifying human papillomavirus type 16 DNA in cervical scrape specimens

Abstract

We compared real-time LightCycler and TaqMan assays and the GP5+/6+ PCR/enzyme immunoassay (EIA) to assess the human papillomavirus type 16 (HPV16) load in cervical scrape specimens. Both real-time PCR assays determined the HPV16 load in scrape specimens similarly. The level of agreement between these assays and the GP5+/6+ PCR/EIA was low (P = 0.004), suggesting that the latter method is not suited for quantifying HPV16 DNA.

FIG. 1.

Correlation of HPV16 DNA loads as determined by different assays. (A) Correlation of TaqMan (y axis) and LightCycler (x axis) assays stratified for cellular input. (B) Correlation of TaqMan (y axis) and LightCycler (x axis) assays in assessing HPV16 load per scrape specimen. (C) GP5+/6+ PCR/EIA OD values (x axis) plotted against the LightCycler HPV16 load/scrape specimen (y axis). (D) GP5+/6+ PCR/EIA OD values (x axis) plotted against the TaqMan HPV16 load/scrape specimen (y axis). All values determined by real-time PCR are ln normalized in the plots. The inner line in each plot represents the regression line, whereas the outer lines indicate the 99% confidence interval of the regression line. Spearman correlation coefficients (ρ) are indicated in each plot.