I-Human Papillomavirus (Iintlobo ezingama-28) iKit yokuFumana iGenotyping (Fluorescence PCR)

Ikiti isebenzisa i-multiple nucleic acid amplification (PCR) indlela yokufumanisa i-fluorescence.Iiprimers ezikhethekileyo kunye neeprobe ziyilwe ngokusekelwe kuhlobo lwe-L1 ekujoliswe kuyo ngokulandelelana kwe-HPV.Iprobe ethile ibhalwe ngeFAM fluorophore (HPV6, 16, 26, 40, 53, 58, 73), VIC/HEX fluorophore (HPV11, 18, 33, 43, 51, 59, 81), CY5 fluorophore, HP435 , 45, 54, 56, 68, 82) kunye ne-ROX fluorophore (HPV31, 39, 42, 52, 61, 66, 83) kwi-5 ', kunye neqela le-3' quencher yiBHQ1 okanye iBHQ2.Ngexesha lokwandiswa kwe-PCR, iiprimers ezithile kunye neeprobes zibophelela kulandelelwano lweethagethi zazo.Xa i-enzyme ye-Taq idibana ne-probes ebophelelwe kulandelelwano olujoliswe kuyo, yenza umsebenzi we-5 'end exonuclease ukwahlula intatheli ye-fluorophore kwi-quencher fluorophore, ukwenzela ukuba inkqubo yokubeka iliso ye-fluorescence inokufumana isignali ye-fluorescent, oko kukuthi, rhoqo xa i-DNA. i-strand ikhulisiwe, i-molecule ye-fluorescent yenziwe, eqonda ulungelelwaniso olupheleleyo lokuqokelelwa kweempawu ze-fluorescent kunye nokwakhiwa kweemveliso ze-PCR, ukuze kuphunyezwe ukuxilongwa kwekhwalithi kunye ne-genotyping ye-nucleic acids ze-28 iintlobo ze-papillomavirus yabantu kwiisampuli zeseli ezikhutshwe emlonyeni wesibeleko. .