title: Real-time duplex PCR for simultaneous HPV 16 and HPV 18 DNA quantitation.
abstract: HPV 16 and HPV 18 are responsible for more than 75% of cervical cancers and high HPV 16 loads are associated with both prevalent and incident lesions. The objective of the present study was to develop a method allowing the detection and quantitation of HPV 16 and 18 DNA to improve future strategies for cervical cancer screening. A duplex real-time PCR allowing the simultaneous quantitation of both HPV 16 and HPV 18 was carried out. Mixes of HPV 16 and HPV 18 whole genome plasmids were prepared to test a wide range of viral DNA concentrations. The values obtained for each mix of plasmids with the simplex and the duplex PCR were very close to the theoretical values except when a HPV type represented only 1:1000 genome equivalent or lower than the concurrent type. Cervical samples harboring HPV 16, HPV 18 or both types were tested by comparing the results with simplex and duplex real-time PCR assays. HPV 16 and HPV 18 genome titers were similar with the two assays. In conclusion, the real-time duplex PCR proved to be robust for HPV 16 and HPV 18 DNA quantitation.
authors.affiliation: Univ Franche-Comte, F-25000 Besancon, France; EA 3181, FED4234, CIC-BT 506, F-25000 Besancon, France.
authors.forename: Elise
authors.initials: E
authors.lastname: Jacquin
authors.forename: Maëlle
authors.initials: M
authors.lastname: Saunier
authors.forename: Frédéric
authors.initials: F
authors.lastname: Mauny
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Schwarz
authors.forename: Christiane
authors.initials: C
authors.lastname: Mougin
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Prétet
mesh_terms: D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D052162:Human papillomavirus 16 / Q000235:genetics / Q000302:isolation & purification*; D052161:Human papillomavirus 18 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D019562:Viral Load / Q000379:methods*
chemical_list: D004279:DNA, Viral
keywords: DNA quantitation; Duplex real-time PCR; HPV 16; HPV 18