title: Quantification of human papilloma virus (HPV) DNA using the Cobas 4800 system in women with and without pathological alterations attributable to the virus.
abstract: OBJECTIVE
  Surrogate markers such as viral load are necessary to follow the evolution of disease resulting from infection with Human Papilloma virus (HPV), especially in this era of vaccination. As such, this paper uses the automated system Cobas-4800-HPV to define viral load as number of HPV copies/cell and apply the results to clinical samples.
  
  
  METHODS
  A curve to determine viral load per cell was constructed from HPV plasmid and cell concentrations using the Cobas-4800-HPV system. According to these curves, HPV viral load was determined in 309 positive endocervical swabs (58 from patients with previous HPV-infection, 118 with current lesions and 133 symptom-free patients presenting for screening) from women attending gynaecology consultations from January to June 2013.
  
  
  RESULTS
  In curves with r(2)≥0.95 the Cobas-4800-HPV system has a detection limit of 150 (2.18 log) viral copies, and the limit for β-globin corresponds to that of a single cell. In women reporting for screening, viral load was under 10(4) (4 log) copies/10(3) cells. For women with lesions or previous HPV infection loads were significantly higher particularly in the 30-45 year group (p=0.038). Elevated viral loads were especially noticeable in non-HPV 16/HPV 18.
  
  
  CONCLUSIONS
  Automated system Cobas-4800-HPV is suitable for define viral load of HPV. Correlation between viral load and number of cells established. Higher viral load in women with disease, and those between 30 and 45 years. Increased viral load of non-16/18 high-risk HPV genotypes detected in patients with lesions compared to screening patients. A difference not observed for HPV 16/18, or in coinfections.
authors.affiliation: Department of Microbiology, Hospital Universitario Central de Asturias, Oviedo, Spain. Electronic address: martaealvarez@gmail.com.
authors.forename: Marta Elena
authors.initials: ME
authors.lastname: Álvarez-Argüelles
authors.affiliation: Department of Microbiology, Hospital Universitario Central de Asturias, Oviedo, Spain. Electronic address: maria.ona@sespa.princast.es.
authors.forename: María
authors.initials: M
authors.lastname: de Oña-Navarro
authors.affiliation: Department of Microbiology, Hospital Universitario Central de Asturias, Oviedo, Spain. Electronic address: ssnrj4@gmail.com.
authors.forename: Susana
authors.initials: S
authors.lastname: Rojo-Alba
authors.affiliation: Department of Gynecology, Hospital Universitario Central de Asturias, Oviedo, Spain. Electronic address: magdalena.torrens@sespa.princast.es.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Torrens-Muns
authors.affiliation: Sexually Transmitted Infections Unit, Hospital Monte Naranco, Oviedo, Spain. Electronic address: marisajunquera@gmail.com.
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Junquera-Llaneza
authors.affiliation: Department of Microbiology, Hospital Universitario Central de Asturias, Oviedo, Spain. Electronic address: jose.boga@sespa.princast.es.
authors.forename: Jose
authors.initials: J
authors.lastname: Antonio-Boga
authors.affiliation: Department of Microbiology, Complejo Hospitalario Universitario de Vigo, Spain. Electronic address: sonia.perez.castro@gmail.com.
authors.forename: Sonia
authors.initials: S
authors.lastname: Pérez-Castro
authors.affiliation: Department of Microbiology, Hospital Universitario Central de Asturias, Oviedo, Spain. Electronic address: santiago.melon@sespa.princast.es.
authors.forename: Santiago
authors.initials: S
authors.lastname: Melón-García
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D057205:Automation, Laboratory / Q000379:methods*; D015331:Cohort Studies; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D005260:Female; D006801:Humans; D008875:Middle Aged; D027383:Papillomaviridae / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000821:virology*; D019562:Viral Load / Q000379:methods*; D055815:Young Adult
chemical_list: D004279:DNA, Viral
keywords: HPV; Intraepithelial lesion; Neoplasic progression; Viral load