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Journal of Clinical Microbiology, September 2006, p. 3292-3298, Vol. 44, No. 9
0095-1137/06/$08.00+0     doi:10.1128/JCM.00539-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Highly Effective Detection of Human Papillomavirus 16 and 18 DNA by a Testing Algorithm Combining Broad-Spectrum and Type-Specific PCR

DDL Diagnostic Laboratory, Fonteijnenburghlaan 5, Voorburg, The Netherlands,¹ GSK Biologicals, Rue de l'Institut 89, Rixensart, Belgium²

Received 13 March 2006/ Returned for modification 8 June 2006/ Accepted 19 June 2006

The use of a single broad-spectrum human papillomavirus (HPV) DNA-based PCR test may fail to detect lower concentrations of HPV DNA due to competition between different genotypes in mixed infections. To improve HPV detection by PCR, broad-spectrum and type-specific (TS) PCRs were combined, with a focus on HPV-16 and HPV-18. Cervical and cervicovaginal cell samples were obtained from 1,113 healthy women (age range, 15 to 25 years) participating in an HPV-16/HPV-18 candidate vaccine efficacy trial. These samples were tested by a broad-spectrum SPF₁₀ PCR-DNA enzyme immunoassay, followed by a primer SPF₁₀-based line probe assay (SPF₁₀ LiPA), and HPV-16- and HPV-18-TS PCRs. The results for the majority of the HPV-16/18 SPF₁₀ LiPA-positive samples were confirmed by TS-PCR (kappa values, 0.775 for HPV-16 and 0.785 for HPV-18). However, TS PCR revealed additional positive samples among those that contained other HPV genotypes due to competition. Conversely, SPF₁₀ LiPA identified HPV-16 or -18 in samples that remained negative by TS PCR as a result of sampling variation. Analysis of follow-up samples from more than 1,000 women confirmed that the combination of SPF₁₀-LiPA with additional HPV-16- and HPV-18-TS PCR diminishes the rate of false-negative diagnosis. The combination of broad-spectrum and TS PCRs resulted in a novel testing algorithm. This combination of assays is more accurate than either method alone, and the novel algorithm offers a highly accurate and effective method for the analysis of HPV infections.

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