High-throughput genotyping of oncogenic human papilloma viruses with MALDI-TOF mass spectrometry.
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.
Söderlund-Strand
Anna
A
WHO HPV LabNet Global Reference Laboratory and Department of Medical Microbiology, Lund University, Malmö University Hospital, Malmö, Sweden.
Dillner
Joakim
J
Carlson
Joyce
J
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D005260:Female; D005838:Genotype; D006801:Humans; D008875:Middle Aged; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000235:genetics*; D016133:Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D002583:Uterine Cervical Neoplasms / Q000821:virology*; D002578:Uterine Cervical Dysplasia / Q000821:virology*