High-throughput genotyping of oncogenic human papilloma viruses with MALDI-TOF mass spectrometry

Abstract

Background: Human papilloma virus (HPV) is the major cause of cervical cancer. Use of HPV genotyping in cervical screening programs and for monitoring the effectiveness of HPV vaccination programs requires access to economical, high-throughput technology.

Methods: We used the Sequenom MassARRAY platform to develop a high-throughput mass spectrometric (MS) method for detecting 14 specific oncogenic HPV genotypes in multiplex PCR products. We compared results from 532 cervical cell samples to the comparison method, reverse dot blot hybridization (RDBH).

Results: The MS method detected all samples found positive by RDBH. In addition, the MS method identified 5 cases of cervical disease (cervical intraepithelial neoplasia of grade I or higher) that RDBH analysis had missed. Discrepancies in specific genotypes were noted in 20 samples, all positive by MS, with an overall concordance of kappa = 0.945.

Conclusions: The MS high-throughput method, with a processing capacity of 10 x 384 samples within 2 working days and at a consumables cost of about US$2 per sample, performed as well as or better than the comparison method.