Multiplex HPV Genotyping of Cervical Scrapes: Comparison of Reverse Line Blot Versus Luminex Technology

Cervical cancer is the second most common cancer in women worldwide. It is causally related to infections with certain, so-called high-risk human papillomavirus (HPV) types. DNA of the known 15 high-risk HPV types can be detected in virtually all cervical cancers. Persistent infection with high-risk types increases the risk for cervical cancer development. HPV types 16 and 18 are most prevalent in cervical cancer, and HPV 16 may have a higher potential for persistence and transformation. Routine screening for precursor lesions is conducted by Papanicolaou staining of exfoliated cervical cells. Type-specific detection of HPV in such samples, however, can provide important additional information, e.g. for patient follow-up. To date, a variety of HPV DNA detection methods exist, some using general PCR amplification primers to enrich for HPV DNA followed by specification of the HPV type. In our laboratory, HPV genotyping is performed with the well validated GP5+/bioGP6+ primer set for HPV DNA amplification followed by hybridisation of the PCR products to membrane-immobilised type-specific oligonucleotide probes (Reverse Line Blot, RLB). Recently, the adoption of this HPV genotyping system to Luminex bead-based suspension array technology was described by Schmitt et al., showing an excellent agreement and even a higher sensitivity compared to RLB. Here we present a follow-up validation of RLB versus Luminex technology for 15 high-risk types, 3 putative high-risk types and 5 low-risk types in 150 clinical samples to compare the two detection methods for their field applicability. Preliminary data obtained so far confirms the higher sensitivity of bead-based multiplex HPV genotyping.