Performance of Commercial Reverse Line Blot Assays for HPV Genotyping.
WHO Human Papillomavirus Laboratory Network Global Reference Laboratory, Chronic Viral Diseases Branch, Division of High-Consequence Pathogens and, National Center for Emerging and Zoonotic Infectious Diseases Pathology, Centers for Disease Control and Prevention, Atlanta, GA 30333, U.S.A.
The performance of three line blot assays (LBA) - Linear Array HPV genotyping assay (LA; Roche Diagnostics), INNO-LiPA HPV Genotyping Extra (LiPA; Innogenetics) and the Reverse Hybridization assay (RH; Qiagen) was evaluated using quantitated whole genomic HPV plasmids (types 6, 11, 16, 18, 31, 33, 35, 39, 51, 52, 56, 58, 59, 68b) as well as epidemiologic samples. In a plasmid titration series LiPA and RH did not detect 50 international units (IU) of HPV 18 in the presence of 5 × 10(4) or more IU HPV16. HPV DNA (1 - 6 types) in the plasmid challenges at 50 (IU) or genome equivalents (GE) were identified with an accuracy of 99.9% by LA, 97.3% by LiPA and 95.4% by RH with positive reproducibility of 99.8% (Kappa = 0.992), 88.2% (Kappa = 0.928) and 88.1% (Kappa = 0.926) respectively. Two instances of mis-typing occured with LiPA. Of the 120 epidemiologic samples 76 were positive for high-risk types by LA, 90 by LiPA and 69 by RH with a positive reproducibility of 87.3% (Kappa = 0.925), 83.9% (Kappa = 0.899) and 90.2% (Kappa = 0.942) respectively. Although all the assays had good concordance in the clinical samples, the greater accuracy and specificity in the plasmid panel suggest that the LA assay has an advantage for internationally comparable genotyping studies.
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