Hybridization assay performed at ambient temperature for typing high-risk human papillomaviruses

BackgroundHuman papillomavirus (HPV) infection with oncogenic types is a prerequisite for cervical cancer development. HPV typing is required in the management of pre-cancerous lesions, epidemiological studies, and vaccination trials. None of the available HPV assays are satisfactory for routine diagnosis.ObjectivesIn order to develop an assay for clinically relevant HPV types, we generated HPV probes using in vitro selection scheme of iterative hybridization.Study designStarting from a mixture of random oligonucleotides, through several rounds of hybridization with 39 type-specific GP5+/6+ L1 sequences, we aimed to obtain specific probes to discriminate between these HPV types.ResultsIn vitro selection led to pools of specific probes, from which individual probes were cloned and tested for their diagnostic performance at ambient temperature. Typically, 10-fold stronger hybridization signals were obtained between each of the selected probes and their specific targets compared to signals with the remaining 38 HPV types. High sensitivity and specificity of selected probes was demonstrated a series of clinical samples in the hybridization assay.ConclusionsA new panel of probes for detecting HPV types is described. Probes can be adapted for use in a simple clinical setting, or incorporated into different detection systems.