Human papillomavirus detection and typing using a nested-PCR-RFLP assay

background: it is clinically important to detect and type human papillomavirus (hpv) in a sensitive and specific manner. objectives: development of a nested-polymerase chain reaction-restriction fragment length polymorphism (nested-pcr-rflp) assay to detect and type hpv based on the analysis of l1 gene. methods: analysis of published dna sequence of mucosal hpv types to select sequences of new primers. design of an original nested-pcr assay using the new primers pair selected and classical my09/11 primers. hpv detection and typing in cervical samples using the nested-pcr-rflp assay. results: the nested-pcr-rflp assay detected and typed hpv in cervical samples. of the total of 128 clinical samples submitted to simple pcr and nested-pcr for detection of hpv, 37 (28.9%) were positive for the virus by both methods and 25 samples were positive only by nested-pcr (67.5% increase in detection rate compared with single pcr). all hpv positive samples were effectively typed by rflp assay. conclusion: the method of nested-pcr proved to be an effective diagnostic tool for hpv detection and typing.