Development And Application Of Gene Typing Of HPV Based On The Magnetic Particles

To develop a gene typing method for 24 kinds of HPV that induce different cancer risk level based on magnetic particle.Through designing specific primer probes,establish a method for detecting the 24 kinds genotype of HPV based on magnetic beads nucleic acid extraction and multiple platform of real-time fluorescent quantitative PCR,by which we determined the reference value of control sample and evaluated its performance.Our method can efficiently genotype the 24 kinds of HPV,the sensitivity is not lower than 100 copies/ml,and the detection range is1.0×10~2~1.0×10~9 copies/ml.The degree of preciseness is higher than 95%in between run and within run.Herpes zoster virus,herpes simplex virus,candida albicans,E.coli,mycoplasma,M.genitalium,mycoplasma,chlamydia trachomatis,neisseria gonorrhea bacteria are no cross reaction with HPV.Urea(6%),inorganic salt(6%),immunoglobulin(1.6 g/dL),white blood cell content(1×10~7/ml),and antibiotics(6 mg/ml)do not affect the HPV detection.Our method can realize 24 kinds of HPV genotypes,is rapid,sensitive screening in one time,and can be used for early diagnosis of cervical cancer.