EVALUATION OF DIFFERENT TECHNIQUES FOR THE IDENTIFICATION OF HUMAN PAPILLOMAVIRUS (HPV) TYPES OF LOW PREVALENCE

Ivan Sabol, Martina Salakova, Jana Smahelova, Michal Pawlita, Markus Schmitt, Nina Milutin Gaperov, Magdalena Grce^*, and Ruth Tachezy

Division of Molecular Medicine, Ruðer Bokovi Institute, Bijenicka 54, 10002 Zagreb, Croatia; Department of Experimental Virology, Institute of Hematology and Blood Transfusion, U Nemocnice 1, 128 20 Prague 2, Czech Republic; Research Program Infection and Cancer, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany

^* To whom correspondence should be addressed. Email: grce{at}irb.hr.

	Abstract

Human papillomaviruses (HPV) have been recognized as an etiologic factor in a variety of diseases. Due to the large number of HPV types, methods for HPV genotyping are difficult to standardize. Despite this fact, several methods exist and some of them are available commercially. In this study, we evaluated the Roche Diagnostics Linear Array (LA) HPV genotyping assay, the Innogenetics INNO-LiPA (LiPA) assay and two non-commercial Reverse Line Blot (RLB) assays based on either GP5+/6+ primers (GP) or newly designed Broad-Spectrum Primers (BS). The reliability of the above-mentioned assays was tested on a wide spectrum of less prevalent HPV types in cervical samples. This is the first study to compare the performance of the most widely used HPV genotyping methods on selected samples positive for low prevalent HPV types. We focused on inter-assay agreement, both overall and type-specific, in cases with single and/or multiple HPV infection. Inter-assay agreement was moderate in single HPV infections and poor in multiple HPV infections. LA and BS-RLB found a higher rate of cases positive for multiple HPV types than LiPA and GP-RLB. The lowest capability in detecting multiple HPV infections was observed for the LiPA assay. The use of only one assay in epidemiological and clinical studies might lead to biased conclusions. Therefore, a universally evaluated and agreed upon HPV typing assay is needed for possible clinical applications, or a combination of current assays, with a knowledge of their limitations, is advised.