Papillomavirus Kanaka (28 ʻAno) ʻĀpana ʻike Genotyping (Fluorescence PCR)

Hoʻohana ka pahu i ke ʻano ʻike fluorescence hoʻonui nui nucleic acid amplification (PCR).Hoʻolālā ʻia nā kumu mua a me nā ʻimi noiʻi ma muli o ka L1 gene target sequence o HPV.Hoʻopili ʻia ka ʻimi kikoʻī me FAM fluorophore (HPV6, 16, 26, 40, 53, 58, 73), VIC/HEX fluorophore (HPV11, 18, 33, 43, 51, 59, 81), CY5 fluorophore (HPV35, 44). , 45, 54, 56, 68, 82) a me ROX fluorophore (HPV31, 39, 42, 52, 61, 66, 83) ma 5', a ʻo ka hui quencher 3 ʻo BHQ1 a i ʻole BHQ2.I ka wā o ka hoʻonui ʻana i ka PCR, hoʻopaʻa ʻia nā kumu mua a me nā probes i kā lākou mau kaʻina pahuhopu.Ke hālāwai ʻo Taq enzyme i nā probes i hoʻopaʻa ʻia i ke kaʻina pahuhopu, hoʻohana ia i ka hana o 5' end exonuclease e hoʻokaʻawale i ka fluorophore mea hoʻopuka mai ka fluorophore quencher, i hiki i ka ʻōnaehana nānā fluorescence ke loaʻa ka hōʻailona fluorescent, ʻo ia hoʻi, i kēlā me kēia manawa he DNA. Hoʻonui ʻia ka strand, hoʻokumu ʻia kahi mole fluorescent, ka mea e ʻike ai i ka hoʻonohonoho piha ʻana o ka hōʻiliʻili ʻana o nā hōʻailona fluorescent a me ka hoʻokumu ʻana o nā huahana PCR, i mea e hoʻokō ai i ka qualitative a me ka genotyping detection o nā nucleic acid o 28 mau ʻano o ka papillomavirus kanaka i loko o nā cervical exfoliated cell samples. .