Detection of high oncogenic-risk papilloma virus in high grade cervical lesions and cancers by a PCR/ELISA assay

摘要

This invention provides a method of detecting a high oncogenic- risk type human papillomavirus in a subject which comprises: obtaining from a subject a specimen containing cervical cells and treating the specimen so as to recover nucleic acid molecules present in the cervical cells; contacting the resulting nucleic acid molecules with multiple pairs of single-stranded labeled oligonucleotide primers capable of specifically hybridizing with a different high oncogenic-risk type of human papillomavirus; amplifying any nucleic acid molecules to which a pair of primers hybridizes so as to obtain a double-stranded amplification product and treating any double-stranded amplification product so as to obtain single-stranded nucleic acid molecules; contacting any resulting single-stranded nucleic acid molecules with multiple single-stranded labeled oligonucleotide probes which are capable of specifically hybridizing with such high oncogenic-risk types of human papillomavirus; contacting any resulting hybrids with a marked antibody capable of specifically forming a complex with the labeled probe, when the probe is present in such a complex; and detecting the presence of any resulting complexes, the presence thereof being indicative of the presence of a high oncogenic-risk type human papillomavirus in the initial specimen.