Clinical Effectiveness of Restriction Fragment Mass Polymorphism Assay for Human Papillomavirus Genotyping

Lee, Eun Hee; Lee, Kyu Bum; Kim, Eun Ok; Ji, Seung Il; Shin, Soo Kyung; Lee, Han Seong; Shim, Soo Yeon; Kim, Soo Ok; Hong, Sun Pyo

Abstract

BACKGROUND
Human papillomavirus (HPV) infection is the main cause of cervical cancer and with the advent of genotype specific vaccines, there is increased need for accurate, broad-spectrum and high-throughput methods for HPV genotyping. A MALDI-TOF mass spectrometry (MS)-based restriction fragment mass polymorphism (RFMP) assay has proven to accurately and reliably genotype a wide variety of HPV.
METHODS
We evaluated the clinical utility of the RFMP assay in HPV genotyping by testing a total of 2,689 specimens taken from liquid-based cytology, which was composed of normal cytology, atypical squamous cells of undetermined significance (ASCUS), low grade squamous intraepithelial lesion (LSIL), high grade squamous intraepithelial lesion (HSIL) and invasive squamous cervical cancer (SCC).
RESULTS
Overall HPV positive rate of total specimens was 32.5% and the high-risk positivity was 16.4%. The HPV positive rates were increased as increasing severity level of cervical lesion. Predominant high-risk HPV genotypes were found as following order; 52 (18.6%), 16 (13.7%), 18 (3.8%), 58 (3.4%), 56 (2.6%) and 31 (2.5%). The high-risk HPV positivities according to cytologic diagnosis were 10.7% (238/2229), 31.7% (76/240), 50.0% (88/176), 86.0% (37/43), 100% (1/1) in normal, ASCUS, LSIL, HSIL and SCC subgroups, respectively. The concordance rate and Kappa value between sequencing and RFMP assays were 96.6% and 0.932 (95%CI: 0.908-0.956).
CONCLUSIONS
The RFMP HPV genotyping assays showed high concordance with sequencing. The assay is simple, and can accurately detect and identify HPV genotypes in samples with various levels of cytological lesions. The results demonstrated that RFMP assay should be clinically suitable for HPV genotyping in laboratories.