title: Development and Validation of a Preanalytic Procedure for Performing the cobas HPV Test in SurePath Preservative Fluid.
abstract: The formation of chemical cross-links between nucleic acids and proteins in formalin-containing media presents challenges for human papillomavirus (HPV) testing of cervical samples collected in SurePath Preservative Fluid. A preanalytic process involving addition of a nucleophilic buffer and heating the sample to 120°C was developed to reverse the effects of cross-linking and improve nucleic acid accessibility for the cobas HPV Test in SurePath. Cycle threshold (CT) values for cobas HPV detection were evaluated over time and various temperatures, and mean CTdifferences between pretreated and both untreated SurePath samples and those collected in PreservCyt were assessed. Without pretreatment, low viral levels (1 × limit of detection) of HPV were no longer detectable by 7 days. For prospectively collected specimens, mean (95% CI) CTdifferences between pretreated and untreated samples indicated enhanced HPV DNA recovery in all categories of treated samples: -2.58 (-3.16 to -2.01), -2.63 (-3.62 to -1.64), and -3.39 (-4.95 to -1.82), respectively, for other 12 high-risk HPV types, HPV16, and HPV18. Furthermore, mean (95% CI) CTdifferences of pretreated SurePath samples were comparable to simultaneously collected PreservCyt samples: -0.48 (-0.98 to 0.02) and -0.23 (-0.93 to 0.46), respectively, for HPV16 and HPV18; a borderline significant difference [-0.35 (-0.57 to -0.13)] was observed for other 12 high-risk HPV types. This preanalytic procedure therefore ensures a validated, safe, and accurate method for cobas HPV testing in SurePath.
authors.affiliation: Roche Molecular Systems, Pleasanton, California. Electronic address: mark.krevolin@roche.com.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Krevolin
authors.affiliation: Roche Molecular Systems, Pleasanton, California.
authors.forename: David
authors.initials: D
authors.lastname: Hardy
authors.affiliation: Roche Molecular Systems, Pleasanton, California.
authors.forename: Jim
authors.initials: J
authors.lastname: Pane
authors.affiliation: Roche Molecular Systems, Pleasanton, California.
authors.forename: Shagufta
authors.initials: S
authors.lastname: Aslam
authors.affiliation: Roche Molecular Systems, Pleasanton, California.
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Behrens
mesh_terms: D000328:Adult; D002584:Cervix Uteri / Q000821:virology; D004279:DNA, Viral / Q000235:genetics; D045604:Extracellular Fluid / Q000821:virology*; D005260:Female; D006801:Humans; D008875:Middle Aged; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods* / Q000592:standards*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods / Q000592:standards; D055815:Young Adult
chemical_list: D004279:DNA, Viral