Comparison of HPV genotyping and methylated ZNF582 as triage for women with equivocal liquid-based cytology results.

Yu-Ligh Liou,Huei-Jen Wang,Shu-Yi Lin,Chi-Feng Chang,Tao-Lan Zhang,Chong-Zhen Qin,Lanqin Cao,Tang-Yuan Chu,Yi Zhang,Yu Zhang,Hong-Hao Zhou,Yingzi Liu

doi:10.1186/s13148-015-0084-2

Abstract

IntroductionThe interpretation of equivocal Papanicolaou (Pap) smear results remains challenging, even with the addition of the high-risk human papillomavirus test (HPV-HR). Recently, methylated zinc finger protein 582 (ZNF582) (ZNF582m) was reported to be highly associated with cervical cancer. In this study, we compared the performance of ZNF582m detection and HPV-HR genotyping in the triage of cervical atypical squamous cell of undetermined significance (ASC-US) and atypical squamous cell - cannot exclude a high-grade lesion (ASC-H).Case descriptionTwo hundred and forty-two subjects with equivocal papanicolaou smear (Pap smear) results were recruited in this hospital-based and case-controlled study. The residual cervical cells in liquid-based cytological test (LBC) containers were used for genomic DNA extraction and then for ZNF582m and HPV-HR detection. The level of ZNF582m was quantified by real-time methylation-specific PCR after bisulfite conversion. The HPV-HR test was performed by using a nested multiplex PCR (NMPCR) assay that combines degenerate E6/E7 consensus primers and HPV type-specific primers.Discussion and evaluationSignificant associations were observed between ZNF582m and the risk of cervical intraepithelial neoplasia grade 3 or higher (CIN3+; odds ratio = 15.52, 95% confidence interval (CI): 7.73 to 31.18). The sensitivity and specificity of ZNF582m for women with CIN3+ were 82.43% and 76.79%, respectively. High sensitivity (99.33%) but low specificity (38.76%) was observed for HPV-HR. When combining both positive results of ZNF582m and HPV-HR, the sensitivity and specificity were 82.43% and 81.55%, respectively. The sensitivity and specificity of ZNF582m or HPV-16/18 were 89.19% and 70.24%, respectively. However, the sensitivity and specificity of ZNF582m combined with HPV-16/18 (both ZNF582m and HPV-16/18 positive results) were 59.46% and 94.64%, respectively.ConclusionsZNF582m provides a promising triage tool for women with ASC.To effectively manage ASC patients, a new strategy co-testing for ZNF582m and HPV-16/18 genotyping was proposed. This strategy could reduce the number of patients referred for colposcopic examination and thus provide a feasible follow-up solution in the regions where colposcopy is not readily available. This strategy could also prevent women from experiencing unnecessary anxiety caused by HPV-HR.

Highlights

The interpretation of equivocal Papanicolaou (Pap) smear results remains challenging, even with the addition of the high-risk human papillomavirus test (HPV-HR)
When the CIN2 or more severe neoplasms were targeted, the sensitivity and accuracy of Methylated ZNF582 (ZNF582m) was decreased to 67.27% and 74.38%, respectively, but the specificity was increased to 80.30%
The sensitivity of ZNF582m combined with High-risk human papillomavirus (HPV-HR), HPV-16/18/52/58, or HPV-16/18 was 99.33%, 95.98%, or 89.19%, respectively, but the specificity was increased with values of 34.02%, 48.81%, and 70.24%, respectively

Summary

Introduction

The interpretation of equivocal Papanicolaou (Pap) smear results remains challenging, even with the addition of the high-risk human papillomavirus test (HPV-HR). We compared the performance of ZNF582m detection and HPV-HR genotyping in the triage of cervical atypical squamous cell of undetermined significance (ASC-US) and atypical squamous cell - cannot exclude a high-grade lesion (ASC-H). The residual cervical cells in liquid-based cytological test (LBC) containers were used for genomic DNA extraction and for ZNF582m and HPV-HR detection. In the case of an ASC smear result, a pathologist has to specify the type as either atypical squamous cell of undetermined significance (ASC-US) or cannot exclude a high-grade lesion (ASC-H). The sensitivity of HPV testing is good; the high prevalence of transient HPV infections limits the specificity of this approach, resulting in high number of colposcopic referrals and unnecessary anxiety for women. There is a need for other markers, which have both high sensitivity and specificity [8,9,10]

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Discussion

Conclusion