Ikhithi Yokuthola Izinguquko ze-KRAS 8 (i-Fluorescence PCR)-RUO
Igama lomkhiqizo
Ikhithi Yokuthola Izinguquko ze-KRAS 8 (i-Fluorescence PCR)-RUO
I-Epidemiology
Ukuguqulwa kwamaphuzu esakhiweni se-KRAS kutholakale ezinhlotsheni eziningana zezimila zabantu, cishe izinga lokuguquka elingu-17% ~ 25% esimila, izinga lokuguquka elingu-15% ~ 30% ezigulini ezinomdlavuza wamaphaphu, izinga lokuguquka elingu-20% ~ 50% ezigulini ezinomdlavuza we-colorectal. Ngenxa yokuthi iphrotheni ye-P21 ebhalwe yi-gene ye-K-ras itholakala phansi kwendlela yokubonisa i-EGFR, ngemva kokuguqulwa kwezakhi zofuzo ze-K-ras, indlela yokubonisa engezansi ihlala isebenza futhi ayithintwa yimithi eqondiswe phezulu ku-EGFR, okuholela ekukhuleni okuqhubekayo kwamaseli anonya. Ukuguqulwa kwezakhi zofuzo ze-K-ras ngokuvamile kunikeza ukumelana nama-inhibitor e-EGFR tyrosine kinase ezigulini ezinomdlavuza wamaphaphu kanye nokumelana nemithi yokulwa nama-anti-EGFR ezigulini ezinomdlavuza we-colorectal [1, 2, 3]. Ngo-2008, i-National Comprehensive Cancer Network (NCCN) yakhipha isiqondiso somtholampilo somdlavuza we-colorectal, esaveza ukuthi izindawo zokuguquka kwezakhi zofuzo ezibangela ukuba i-K-ras isebenze zitholakala kakhulu kuma-codon 12 no-13 e-exon 2, futhi yancoma ukuthi zonke iziguli ezinomdlavuza we-colorectal osakazeke kakhulu zingahlolwa ukuze zihlolwe ukuguquka kwe-K-ras ngaphambi kokwelashwa [4]. Ngakho-ke, ukutholakala okusheshayo nokunembile kokuguqulwa kwezakhi zofuzo ze-K-ras kubaluleke kakhulu ekuqondisweni kwemithi yokwelapha. Le khithi isebenzisa i-DNA njengesampula yokuthola ukuze inikeze ukuhlolwa kwekhwalithi kwesimo sokuguquka kwezakhi zofuzo, okungasiza odokotela ekuhloleni umdlavuza we-colorectal, umdlavuza wamaphaphu kanye nezinye iziguli ezinezimila ezizuza emithini eqondiwe. Imiphumela yokuhlolwa kwekhithi ingeyokubhekisela emtholampilo kuphela futhi akufanele isetshenziswe njengesisekelo sodwa sokwelashwa kweziguli ngabanye. Odokotela kufanele benze izahlulelo eziphelele ngemiphumela yokuhlolwa ngokusekelwe ezicini ezifana nesimo sesiguli, izinkomba zemithi, impendulo yokwelashwa kanye nezinye izinkomba zokuhlolwa kwelabhorethri.
Amapharamitha Obuchwepheshe
| Isitoreji | ≤-18℃ |
| Isikhathi sokuphelelwa yisikhathi | Izinyanga ezingu-12 |
| Uhlobo Lwesifanekiso | izingxenye zezifo ezifakwe iparafini kubantu |
| CV | ≤5.0% |
| I-LoD | a) I-K-ras Reaction Buffer A kanye ne-K-ras Reaction Buffer B zingakwazi ukubona izinga lokuguquka elingu-1% ngaphansi kwesizinda se-wild-type esingu-3ng/μL; b) Ukuguqulwa okungu-1×103Amakhophi/i-mL angatholakala ngokuzinzile ngemuva kohlobo lwasendle lwe-1×105Amakhophi/mL uma izinga lokuguqulwa kwezakhi zofuzo lingu-1%; c) Uma ireferensi ye-LoD yenkampani i-SW3 ihlolwa, akukho value ye-Ct noma i-Ct value=0 ye-Reaction Buffer A kanye ne-Reaction Buffer B |
| Izinsimbi Ezisebenzayo | Izinhlelo ze-Biosystems ezisetshenzisiwe 7500 Izinhlelo ze-PCR Zesikhathi Sangempela, Izinhlelo ze-Biosystems ezisetshenzisiwe 7300 Izinhlelo ze-PCR Zesikhathi Sangempela, Izinhlelo ze-QuantStudio®5 ze-PCR Zesikhathi Sangempela, Uhlelo lwe-LightCycler® 480 lwe-PCR Yesikhathi Sangempela, Uhlelo lwe-BioRad CFX96 lwe-PCR Yesikhathi Sangempela. |
Ukugeleza Komsebenzi
Kudingeka i-reagent kodwa ayinikezwanga:I-DNase/RNase H2O engenayo i-DNase, i-ethanol engenamanzi. Uma uhlola isampula yezicubu ezifakwe i-paraffin, kunconywa ukusebenzisa i-QIAGEN's QIAamp DNA FFPE Tissue Kit (56404) kanye ne-Paraffin-embedded Tissue DNA Rapid Extraction Kit (DP330) eyenziwe yi-Tiangen Biotech(Beijing) Co., Ltd.
Izinto ezisetshenziswayo ziyadingeka kodwa azinikezwanga:Amathiphu angenazo i-DNase/RNase, amagilavu angasetshenziswa, ishubhu le-Centrifuge elingenazo i-DNase/RNase, imichilo yamashubhu angu-8, i-centrifuge.
