Amplification with molecular beacon primers and reverse line blotting for the detection and typing of human papillomaviruses

A novel method for the detection and typing of human papillomavirus (HPV) w as developed using molecular beacon primers. The method is based on the use of HPV-specific primers containing a hairpin loop structure in which fluor escent donor and quencher groups are held in close proximity such that fluo rescence is quenched. Amplification of the target sequence results in the o pening of the loop and the resulting fluorescence can be detected on a sequ ence detector system (SDS) 7700 (Applied Biosystems), as used for TaqMan(TM ) assays. Fluorescent amplicons were identified on the SDS 7700 and then ty ped by a single hybridisation with specific probes immobilised in lines on a nylon membrane and detected on a fluorescent scanner. This novel beacon p rimer method compared well with conventional PCR for cervical scrape specim ens. The combination of the beacon primer method and reverse line blotting should enable large-scale population studies of HPV infection. (C) 2000 Els evier Science B.V. All rights reserved.