Effect of Glacial Acetic Acid Treatment of Liquid-Based Cytology Collections on Performance of Cervista HPV HR for Detection of High-Risk Human Papillomavirus

Abstract

cInadequate cervical cytological analysis can be facilitated by glacial acetic acid (GAA) treatment of primary liquid-based collections to remove mucus, erythrocytes, inflammatory cells, and cellular debris. In the context of a commercial human papillomavirus (HPV) hybridization assay performed on 465 tandem specimens with and without GAA treatment, we show that GAA treatment significantly reduces genomic DNA content (P < 0.0001) and creates an increased potential for indeterminate and falsenegative results. In the context of cytological workflow, laboratories should consider providing a specimen aliquot for HPV DNA detection prior to GAA treatment. Molecular diagnostic assays specific for human papillomavirus (HPV) play a significant role in cervical cancer triage (15). Such assays, shown to have Ն98.9% negative predictive value for the underlying presence of cervical intraepithelial neoplasia 2ϩ (5, 13), can be performed on endocervical scrapings preserved in liquid transport medium. This collection, also forwarded for cytological analysis, has the potential to possess insufficient squamous cellularity, as well as abundant erythrocytes, mucus, acute inflammatory cells, and cytological debris (11). In particular, the latter factors may compromise Papanicolaou (Pap) smear analyses that are reliant on filter-based processing protocols (e.g., ThinPrep; Hologic, Marlborough, MA). As a result, glacial acetic acid (GAA)-based protocols have been developed to augment diagnostic yield, with some receiving U.S. Food and Drug Administration approval (4).While this paradigm has procured a high percentage of newly satisfactory Pap smear analyses and has increased detection of significant gynecological lesions (1, 2, 6, 11), recent data have revealed that GAA treatment increases false-positive diagnoses of atypical endocervical cells (3). A few studies, typically focused on hybrid capture technology (H c 2; Qiagen/Digene, Gaithersburg, MD), have been conflicting on the potential effect of GAA on HPV DNA detection (1, 6). In addition to oligonucleotide probes specific for 14 high-risk HPV types (provided in three reagent mixtures), the Cervista HPV HR (Hologic/Third Wave Technologies, Madison, WI) possesses an oligonucleotide probe to assess relative DNA content. The presented study utilizes this internal control as one means of assessing interference potential of GAA treatment on assay performance.In an institutional review board-approved protocol, 465 ThinPrep specimens were collected by two laboratories. These specimens were initially defined as unsatisfactory on the basis of the 2001 Bethesda system guidelines (12) for assessing quality of Pap smears. Laboratory A subjected unsatisfactory ThinPrep specimens to a manufacturer-advocated protocol in which unsatisfactory specimens were initially treated with a titration of GAA and CytoLyt solution (Hologic) (2, 4). In contrast, laboratory B validated a protocol derived from the reports of Iverson and Armour (8) and Agoff et al. (1) in which concentrated GAA was added to spec...

Comparative study of the cervista and hybrid capture 2 methods in detecting high‐risk human papillomavirus in cervical lesions

“…The Cervista HPV HR test is a second method approved by the US FDA for HPV DNA detection. Because of relatively short periods of approval, there are few studies on the clinical utilization of the Cervista HPV HR test . In this study, we compared the two HPV testing methods with regard to HR HPV detection rate, triage of ASCUS, and prediction of cervical disease progression.…”

Effect of Preanalytical Processing of ThinPrep Specimens on Detection of High-Risk Human Papillomavirus by the Aptima HPV Assay

“…We previously demonstrated that Cervista is greatly affected by GAA treatment (10). Analysis of 465 specimens revealed significant decreases in gDNA content and rates of positive HPV results upon GAA treatment.…”

“…In addition, specimen carryover (potentiated by either form of automation) may be of concern to both laboratorians and cytopathologists. Moreover, previous data (8)(9)(10) suggest that performance characteristics of DNA-based HPV modalities may be affected by glacial acetic acid (GAA) treatment of liquid-based cytology collections. The purpose of this investigation was to assess such preanalytical concerns inherent to the performance of the Aptima HPV.…”

Clinical sensitivity of HPV assays for the detection of high grade cervical disease in cervical samples treated with glacial acetic acid

“…Of the small number of studies where the impact of GAA has been assessed, the evidence would indicate that the COBAS 4800HPV Assay (Roche Molecular Systems, Pleasanton, CA, USA) and APTIMA HPV Test (Gen-Probe Inc., San Diego, CA) are unaffected by GAA treatment − as opposed to the Cervista Test (Hologic, Inc., Bedford, MA) where it has a deleterious influence [3][4][5][6]. Furthermore, in an earlier splitspecimen study of untreated vs treated specimens undertaken in our laboratory, while the impact of GAA on HPV detection was insignificant at the qualitative level, GAA treatment was associated with lower read-outs according to the semi-quantitative measures of the assays chosen: HC2 and rtHPV [7].…”