I-Real-time PCR Systems: Ithuthukisa Ucwaningo Nokuxilongwa

Amasistimu we-PCR wesikhathi sangempelabaye bashintsha imikhakha yebhayoloji yamangqamuzana nokuxilonga ngokunikeza abacwaningi nodokotela amathuluzi anamandla okuhlaziya ama-nucleic acid. Ubuchwepheshe bungathola futhi bulinganise ukulandelana kwe-DNA ethile noma i-RNA ngesikhathi sangempela, kuyenze ibe yimpahla ebalulekile ocwaningweni oluhlukahlukene kanye nasezinhlelweni zokuxilonga.

Enye yezinzuzo eziyinhloko zezinhlelo ze-PCR zesikhathi sangempela yikhono lazo lokuhlinzeka ngemiphumela esheshayo, nenembile. Izindlela zendabuko ze-PCR zidinga ukuhlaziywa kwangemuva kokukhulisa, okungadla isikhathi futhi kube nzima. Ngokuphambene, izinhlelo ze-PCR zesikhathi sangempela zivumela abacwaningi ukuthi baqaphe ukukhuliswa kwe-DNA noma i-RNA, ngaleyo ndlela bathole ukulandelana okuqondiwe ngesikhathi sangempela. Lokhu akugcini nje kongi isikhathi kodwa futhi kunciphisa ubungozi bokungcola kanye namaphutha omuntu, okwenza i-PCR yesikhathi sangempela ibe ubuchwepheshe bokuhlaziya amangqamuzana obusebenzayo nobuthembekile.

Ezilungiselelweni zocwaningo, amasistimu e-PCR esikhathi sangempela asetshenziswa kakhulu ekuhlaziyeni isisho sezakhi zofuzo, i-genotyping, kanye nokutholwa kwe-microbial. Ikhono lokulinganisa amazinga wokuvezwa kofuzo ngesikhathi sangempela lithuthukise kakhulu ukuqonda kwethu izinqubo ezihlukahlukene zebhayoloji nezindlela zezifo. Abacwaningi bangasebenzisa i-PCR yesikhathi sangempela ukutadisha imiphumela yokwelashwa okuhlukile noma izimo ekukhulumeni kwezakhi zofuzo, banikeze imininingwane ebalulekile ngesisekelo samangqamuzana esifo kanye nezinhloso zokwelashwa ezingaba khona.

Izinhlelo ze-PCR zesikhathi sangempela nazo ziwusizo ocwaningweni lwe-genotyping ukuhlonza ngokushesha nangokunembile okuhlukile kofuzo kanye nama-polymorphisms. Lokhu kubaluleke kakhulu ezindaweni ezifana ne-pharmacogenomics kanye nemithi yomuntu siqu, lapho ukuhluka kofuzo kungase kube nomthelela ekuphenduleni komuntu emithini nasezinhlotsheni zokwelapha. Ngokusebenzisa ubuchwepheshe be-PCR besikhathi sangempela, abacwaningi bangakwazi ukuhlola ngokuphumelelayo izimpawu zofuzo ezihlobene ne-metabolism yezidakamizwa, ukuthambekela kwezifo, kanye nemiphumela yokwelashwa.

Emkhakheni wokuxilongwa, amasistimu e-PCR esikhathi sangempela adlala indima ebalulekile ekutholeni nasekuqaphelweni kwezifo ezithathelwanayo, izifo zofuzo, nomdlavuza. Ukuzwela okuphezulu kanye nokucaciswa kwe-PCR yesikhathi sangempela kuyenza inkundla ekahle yokuhlonza amagciwane afana namagciwane namagciwane kumasampula omtholampilo. Lokhu kubaluleke kakhulu ophenyweni lokuqubuka kwezifo kanye nemizamo yokuqapha, lapho ukutholakala ngesikhathi nenembile kwemithombo yokutheleleka kubalulekile ekungeneleleni kwezempilo komphakathi.

Ngaphezu kwalokho, izinhlelo ze-PCR zesikhathi sangempela zisetshenziswa kabanzi ekuxilongeni nasekuqapheni izifo zofuzo nomdlavuza. Ngokuqondisa ukuguqulwa kwezakhi zofuzo ezithile noma amaphethini wokuvezwa kwezakhi zofuzo okungajwayelekile, odokotela bangasebenzisa i-PCR yesikhathi sangempela ukuze basize ekutholeni kusenesikhathi, ukubikezela, kanye nokuhlola impendulo yokwelashwa kwezinhlobonhlobo zezifo zofuzo kanye ne-oncological. Thuthukisa ngendlela emangalisayo ukunakekelwa kwesiguli ngokuvumela ukwelashwa komuntu siqu nokuhlosiwe okusekelwe ezicini zamangqamuzana ezifo ngazinye.

Njengoba ubuchwepheshe be-PCR besikhathi sangempela buqhubeka nokuvela, intuthuko entsha efana ne-multiplex PCR kanye ne-PCR yedijithali ithuthukisa futhi amakhono ayo okucwaninga kanye nokuxilonga. I-Multiplex real-time PCR ingathola ukulandelana kwethagethi eminingi ngesikhathi esisodwa ngokusabela okukodwa, yandise ububanzi bokuhlaziya amangqamuzana futhi ilondoloze isampula lezinto ezibalulekile. I-Digital PCR, ngakolunye uhlangothi, ihlinzeka ngenani eliphelele lama-nucleic acid ngokusabalalisa ama-molecule ngamanye ezinkulungwaneni zamakamelo okusabela, anikeze ukuzwela okungenakuqhathaniswa nokunemba.

Ngokufigqiwe,izinhlelo ze-PCR zesikhathi sangempelasebeyithuluzi elibalulekile lokuthuthukisa amakhono okucwaninga kanye nokuxilonga kubhayoloji yamangqamuzana nemithi yokwelapha. Ikhono labo lokuhlinzeka ngokuhlaziywa kwe-nucleic acid esheshayo, enembile, kanye nenani liye lashintsha ukuqonda kwethu izinqubo zezinto eziphilayo kanye nezindlela zezifo futhi lathuthukisa ukuxilonga nokuphathwa kwezimo ezihlukahlukene zezempilo. Njengoba ubuchwepheshe buqhubeka nokusungula, izinhlelo ze-PCR zesikhathi sangempela ze-fluorescence quantitative zizoqhubeka nokukhuthaza ukuthuthukiswa kocwaningo lwesayensi nokunakekelwa kwezokwelapha, ekugcineni kuzuzise iziguli kanye nomphakathi uwonke.