The invention belongs to the technical field of diagnostic reagents, and particularly provides a method for amplifying and
genotyping nucleic acid genes of HPV (
Human Papilloma Virus) and an
assay kit for the same. The method comprises the following steps of: carrying out multiple real-time
quantitative fluorescence gene amplifications (QPCR) by using a
hybrid primer; quantitatively determining high-risk subtype HPVs by using a quantitative probe; and differentiating HPV 16 and 18 subtypes in types by using a
typing probe. The
hybrid primer is designed by conservative sequences at two sides of a special area of an E1
gene encoded by the HPV; and the
genotyping probe is designed according to a type-specific sequence at the center of the area. Gradient dilutions of recombinant plasmids containing 16 and 18 subtype E1-area genes are taken as standard reference. The kit comprises a cervical
brush, a sample
storage tube / liquid, a primer, a probe, a Taq
enzyme & reaction
buffer solution, and a standard contrast reference. The
system can be used for quantitatively detecting 13 high-risk subtype HPVs (HPV-16, HPV-18, HPV-31, HPV-33, HPV-35, HPV-39, HPV-45, HPV-51, HPV-52, HPV-56, HPV-58, HPV-59, HPV-68) and differentiating two subtypes HPV-16 and HPV-18 in types.