title: Human papillomavirus quantification in urine and cervical samples by using the Mx4000 and LightCycler general real-time PCR systems.
abstract: During the last decade, growing efforts have focused on human papillomavirus (HPV) detection using liquid hybridization, conventional PCR, and real-time PCR-based methods to increase the overall proportion of patients participating in cervical cancer screening procedures. We proposed a new general HPV DNA real-time PCR on the Mx4000 (Stratagene) and LightCycler (Roche Diagnostics) systems usable for both cervical scrape specimens and urine samples. A linear range was obtained from 5 DNA copies to 8 log(10) DNA copies/ml, and intra- and interassay variations were between 1.8 and 4%. Cervical carcinoma and HPV DNA screening was performed in 333 individual women referred for gynecological examination at the university hospitals of Angers and Brest and enrolled in the PapU study. Among cervical specimens (n = 333), 45% were positive for HPV DNA, with a mean viral load at 5.00 log/ml (+/- 1.73). Among urine samples (n = 177), 37% were positive with a significant 50-fold-lower mean viral load (3.77 +/- 1.32 log/ml; P < 0.0001). Kappa agreement for HPV DNA between cervical and urine specimens was excellent (93%). Thus, we developed a highly sensitive and quantitative general HPV DNA real-time PCR method that allows mass screening of patients with HPV infection. The ongoing longitudinal and prospective multicenter PapU study should give us the opportunity to validate this method adapted to HPV DNA screening in urine samples in a larger population.
authors.affiliation: Département de Microbiologie-EA3882, CHU Brest, France. christopher.payan@chu-brest.fr
authors.forename: Christopher
authors.initials: C
authors.lastname: Payan
authors.forename: Alexandra
authors.initials: A
authors.lastname: Ducancelle
authors.forename: Mohamed H
authors.initials: MH
authors.lastname: Aboubaker
authors.forename: Julien
authors.initials: J
authors.lastname: Caer
authors.forename: Malena
authors.initials: M
authors.lastname: Tapia
authors.forename: Amelie
authors.initials: A
authors.lastname: Chauvin
authors.forename: Damien
authors.initials: D
authors.lastname: Peyronnet
authors.forename: Elodie
authors.initials: E
authors.lastname: Le Hen
authors.forename: Zohra
authors.initials: Z
authors.lastname: Arab
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Legrand
authors.forename: Adissa
authors.initials: A
authors.lastname: Tran
authors.forename: Edith
authors.initials: E
authors.lastname: Postec
authors.forename: Françoise
authors.initials: F
authors.lastname: Tourmen
authors.forename: Martine
authors.initials: M
authors.lastname: Avenel
authors.forename: Chantal
authors.initials: C
authors.lastname: Malbois
authors.forename: Marie-Anne
authors.initials: MA
authors.lastname: De Brux
authors.forename: Philippe
authors.initials: P
authors.lastname: Descamps
authors.forename: Francoise
authors.initials: F
authors.lastname: Lunel
mesh_terms: D002584:Cervix Uteri / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis / Q000302:isolation & purification; D005260:Female; D006801:Humans; D008403:Mass Screening / Q000379:methods; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014556:Urine / Q000821:virology*; D002583:Uterine Cervical Neoplasms / Q000821:virology
chemical_list: D004279:DNA, Viral