Performance evaluation of Anyplex™II HPV28 detection kit in a routine diagnostic setting: Comparison with the HPV Sign® Genotyping Test

• Published in: Journal of virological methods 2015-06, Vol.217, p.8-13
• Main Authors: Marcuccilli, Fabbio, Farchi, Francesca, Mirandola, Walter, Ciccozzi, Massimo, Paba, Pierpaolo, Bonanno, Elena, Perno, Carlo Federico, Ciotti, Marco
• Format: Article
• Language: English
• Subjects: Cervix Uteri - virology; Female; Genotyping Techniques - methods; HPV; HPV molecular diagnosis; Humans; Multiplex real-time PCR; Papillomaviridae - classification; Papillomaviridae - genetics; Papillomaviridae - isolation & purification; Papillomavirus Infections - diagnosis; Papillomavirus Infections - virology; Sensitivity and Specificity
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2015.02.018

•Anyplex™II HPV28 detection kit is a novel assay for human papillomavirus genotyping based on Tagging Oligonucleotide Cleavage and Extension (TOCE™) technology.•It is a semiquantitative real-time multiplex PCR assay.•It can detect multiple targets in a single fluorescent channel.•It allows simultaneous detection of 19 high-risk HPVs (16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 69, 73, 82) and 9 low-risk HPVs (6,11, 40, 42, 43, 44, 54, 61, 70). Anyplex™II HPV28 is a new PCR assay designed for HPV genotyping. It can detect 28 HPV types including 19 high-risk and 9 low-risk types. This study evaluated the performance of Anyplex™II HPV28 on 123 fresh cervical samples screened in parallel with HPV Sign® Genotyping Test. Of the 123 samples screened, 93 were positive, 15 negative, and 15 discordant. The total number of HPV positive samples combined was 108: 38 single infections and 70 multiple infections. The agreement between the two tests was 87.8%, κ=0.592. Genotype specific agreement was strong for HPV 16 (k=0.761), HPV 18 (k=0.674), and HPV 35 (k=0.796). Sensitivity and specificity of Anyplex™II HPV28 assay using HPV Sign® Genotyping Test as reference was 84.8% and 94%; conversely, sensitivity and specificity of HPV Sign® Genotyping Test was 29% and 99.5%. Anyplex™II HPV28 assay is a sensitive and specific assay suitable for HPV genotyping but requires clinical validation.