title: Analytical performance of RNA isolated from BD SurePath™ cervical cytology specimens by the PreTect™ HPV-Proofer assay.
abstract: Several commercial HPV ancillary tests are available for detection of E6/E7 RNA. It is not clear how storage of a cervical Pap affects the analytical and clinical performance of the PreTect™ HPV-Proofer assay. To investigate the qualitative performance of RNA extracted from BD SurePath™ liquid-based cytology (LBC) specimens for the detection of human papillomavirus (HPV) E6/E7 mRNA using the PreTect™ HPV-Proofer assay, studies including stability, reproducibility, residual specimen analysis, and storage medium comparison assays were performed. Cervical cytology specimens were collected and stored in BD SurePath™ LBC preservative fluid and/or PreTect™ Transport Media. RNA was isolated using the RecoverAll™ Total Nucleic Acid Isolation kit and RNA integrity was evaluated in the PreTect™ HPV-Proofer assay. The performance of RNA isolated from cervical cells collected and stored in BD SurePath™ LBC preservative fluid or PreTect™ Transport Media was also evaluated through a storage medium comparison study. The RNA was found to be stable for a minimum of 21 days when stored at ambient temperature and displayed high reproducibility with the mean percentage reproducibility ranging from 90.5% to 100% for the HPV types detected by the PreTect™ HPV-Proofer assay. The prevalence rate of HPV types in this study cohort was consistent with published reports. A 93.7% first pass acceptance rate was demonstrated across all cytology grades. The positive human U1 snRNP specific A protein (U1A) and HPV rate for BD SurePath™ LBC and PreTect™ Transport Media specimens was statistically equivalent for both normal and abnormal specimens. This data support the use of RNA isolated from BD SurePath™ LBC for ancillary HPV testing and demonstrates the feasibility of using BD SurePath™ preservative fluid as a specimen type with the PreTect™ HPV-Proofer assay.
authors: BD Diagnostics, Women's Health and Cancer, Durham, NC, USA. Eric_Dixon@BD.com
authors: Eric P
authors: EP
authors: Dixon
authors: Petter
authors: P
authors: Grønn
authors: Lorraine M
authors: LM
authors: King
authors: Heather
authors: H
authors: Passineau
authors: Hema
authors: H
authors: Doobay
authors: Hanne
authors: H
authors: Skomedal
authors: Jalil
authors: J
authors: Hariri
authors: Shauna N
authors: SN
authors: Hay
authors: Charlotte A
authors: CA
authors: Brown
authors: Timothy J
authors: TJ
authors: Fischer
authors: Douglas P
authors: DP
authors: Malinowski
mesh_terms: D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics; D005260:Female; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000032:analysis / Q000235:genetics; D027383:Papillomaviridae / Q000302:isolation & purification* / Q000472:pathogenicity; D030361:Papillomavirus Infections / Q000175:diagnosis*; D015995:Prevalence; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D012097:Repressor Proteins / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000032:analysis; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*; D013997:Time Factors; D014626:Vaginal Smears / Q000379:methods
chemical_list: D004268:DNA-Binding Proteins; C068651:E6 protein, Human papillomavirus type 16; C052633:E7 protein, Human papillomavirus type 18; D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic; D012097:Repressor Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear; C075339:U1A protein