Author/Editor		Hošnjak, Lea; Fujs Komloš, Kristina; Kocjan, Boštjan; Seme, Katja; Poljak, Mario
Title		Development of a novel multiplex type-specific quantitative realtime PCR for detection and differentiation of infections with human papillomavirus types HPV2, HPV27, and HPV57
Type		članek
Vol. and No.		Letnik 25, št. 4
Publication year		2016
Volume		str. 65-71
ISSN		1318-4458 - Acta dermatovenerologica Alpina, Panonica, et Adriatica
Language		eng
Abstract		Introduction: The present study describes the development and evaluation of the first multiplex type-specific quantitative realtime PCR (RT-PCR), enabling simple, rapid, sensitive, and specific concurrent detection and differentiation of human papillomavirus (HPV) types HPV2, 27, and 57 in a single PCR reaction. Results: The HPV2/27/57 multiplex RT-PCR with a dynamic range of seven orders of magnitude (discriminating 10 to 10[sup]8 viral genome equivalents/reaction) has an analytical sensitivity of at least 10 viral copies of each targeted HPV type/reaction, and no cross-reactivities were observed among the included targets. All three primer/probe combinations were efficient in amplifying 500 copies of targeted DNA in a background of 10[sup]8, 10[sup]7, 500, 100, and 10 copies of non-targeted viral DNA/reaction, and the performance of the HPV2/27/57 multiplex RT-PCR was additionally not affected by the presence of background human genomic DNA. When testing DNA isolates obtained from fresh-frozen tissue specimens of various children's warts, the results of the HPV2/27/57 multiplex RT-PCR were completely in line with the results of the conventional Low-risk Alpha-PV PCR. Conclusion: The newly developed HPV2/27/57 multiplex RT-PCR is an appropriate test for use in routine clinical laboratory settings and for studies focusing on the molecular epidemiology, pathogenesis, and natural history of HPV2/27/57-related lesions.
Keywords		človeške vrste papiloma virusa odkrivanje diferenciacija human papillomavirus types detection differentiation