Real-time duplex PCR for simultaneous HPV 16 and HPV 18 DNA quantitation.
Fiche publication
Date publication
novembre 2013
Journal
Journal of virological methods
Auteurs
Membres identifiés du Cancéropôle Est :
Pr PRETET Jean-Luc, Pr MAUNY Frédéric
Tous les auteurs :
Jacquin E, Saunier M, Mauny F, Schwarz E, Mougin C, Prétet JL
Lien Pubmed
Résumé
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.
Mots clés
DNA, Viral, genetics, Female, Human papillomavirus 16, genetics, Human papillomavirus 18, genetics, Humans, Multiplex Polymerase Chain Reaction, methods, Papillomavirus Infections, diagnosis, Real-Time Polymerase Chain Reaction, methods, Viral Load, methods
Référence
J. Virol. Methods. 2013 Nov;193(2):498-502