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I-Nucleic acid yokuthola i-fluorescence quantitative value Ct – ipharamitha yenzondo yothando

I-Nucleic acid yokuthola i-fluorescence quantitative value Ct – ipharamitha yenzondo yothando

“Uhlelo Olusha Lokuxilongwa Nokwelashwa Kwe-Coronary Virus Pneumonia (I-Trial Ninth Edition)” lisebenzisa inani le-Ct lokutholwa kwe-nucleic acid ≥35 njengesisekelo esibalulekile sokukhululwa kokulawulwa kokuzihlukanisa noma ukukhishwa.Ngakho, inani le-Ct lekhithi yokuthola i-nucleic acid limelelani?Ingabe amanani e-Ct amakhithi ahlukene ayaqhathaniswa?Ingabe ukuncipha kwevelu ye-Ct, kungcono ukusebenza kwekhithi?
Inani le-Ct (I-Threshold Cycle, Ct) inombolo yemijikelezo ye-PCR lapho ukushuba kwesignali yomthamo we-fluorescence yesikhathi sangempela yeqa umkhawulo omisiwe.Amakhithi okuthola i-Nucleic acid athatha umqhele omusha njengesibonelo.Ngokusabela okufanayo kwekhithi efanayo ukuthola amasampula amabili aqukethe amagciwane, A no-B, usayizi wevelu ye-Ct umele, ngokwezinga elithile, inani lamakhophi ofuzo lwegciwane, okungukuthi, inani legciwane egazini.Uma iphansi i-Ct yesampula B, iyanda inani legciwane egazini.Inani le-Ct lilingana ngokuphambene nomthamo wegciwane egazini kanye nokutheleleka kwalo.Ubuhle nobubi bamakhithi omthamo we-fluorescence wesikhathi sangempela awanakwahlulelwa kuphela ngosayizi wevelu ye-Ct, ngoba amanani e-Ct amakhithi ahlukene awaqhathanisi.Ukuhlolwa kokusebenza kwekhithi kudinga ukulinganiswa ngezici zokuzwela (umkhawulo wokuthola ubuncane), ukucaciswa, ukunemba, ukuzinza, kanye nohlelo lokusebenza lokuxilonga.Umkhiqizi ngamunye we-reagent unezinhlobo zakhe eziyishumi nesishiyagalombili zobuciko bokulwa, futhi banamandla abo omlingo ekusebenziseni, ngakho-ke ngeke ngingene emininingwaneni lapha.
Ngamathuluzi/amasistimu okuhlaziya we-real-time quantitative quantitative PCR (QPCR), ukutholwa kwamanani e-Ct kanye nokusetshenziswa kwawo okusabalele ezinhlelweni zokutholwa kwe-pathogen kujwayelekile kuwo wonke umuntu.Ngokuthuthuka kwemboni kanye nokuthuthuka kobuchwepheshe, imishini ye-QPCR nama-reagents ahlangabezane namathuba nezinselele.I-Yiou Think Tank (Umbiko Wokucwaninga Ka-2021 Ngemboni Yokuhlola Izakhi Zofuzo: Ubuchwepheshe) yase-China ihlaziye ukuthi izinkampani eziningi embonini yezwe lami ye-QPCR zibhekene nezinkinga ezivamile ezisekelwe kubuchwepheshe.Ekuphenduleni lezi zinkinga, izixazululo zihlukaniswa kakhulu zibe izinhlobo ezimbili: ukusheshisa ukuthuthukiswa kobuchwepheshe kanye nokusebenzisa ubuchwepheshe obusha.
I-POCT yokuxilongwa kwamangqamuzana e-QPCR ingenye yezinkomba zokuthuthukiswa kwemboni, futhi ukuthuthukiswa kokusebenza kuyisici esibalulekile sokusheshisa ukusetshenziswa kobuchwepheshe.Emishinini ye-POCT enesampula elilodwa lokuphuma kwe-QPCR, ulikhulisa kanjani inani le-Ct lepharamitha eyinhloko yokusabela okukodwa ku-R&D?Okokusebenza okufanayo, isistimu yokusabela efanayo, kanye nokugxiliswa kwesifanekiso esifanayo, ngokombono likhulu inani le-Ct lokusabela okukodwa, kunciphisa ukusebenza kahle kwe-enzyme ekuphenduleni.
Enqubweni yokuthuthukisa ukusebenza kahle kokusabela kwe-enzyme nokwandisa inani le-Ct, ijika elivamile le-40 PCR lomjikelezo we-fluorescence we-amplification quantitative lingahlaziywa kuqala.Ijika lokukhulisa emfanekisweni ongezansi lihlukaniswe ngesigaba sangemuva se-fluorescence, isigaba sokukhulisa umchazi, isigaba somugqa, kanye nesigaba sethafa.Ukuze uzuze ingemuva le-fluorescence eliphansi, ngokusemandleni akho ukuze lihlukanise inani le-fluorescence lokukhulisa umchazi wokuqala, kuyadingeka ukuhlola kusukela kusithwali sokusabela okusebenzisekayo, ukuhlolwa komklamo we-primer probe, kanye nesistimu yebhafa yokusabela.Isikhathi sokukhulisa i-exponential siwukusabela okuqondile kakhulu komsebenzi we-enzyme, futhi siwubufakazi bokuxhumana okuphelele kwemishini ye-POCT, izithwali zokusabela kanye nama-reagents.Ngalesi sikhathi, ukulawulwa kwezinga lokushisa kwemishini, ukutholwa kwesignali ye-optical kanye nokuhlaziywa, i-biocompatibility yenkampani yenethiwekhi, nokusebenza kwesistimu yama-reagents kudinga ukulungiswa futhi kuhlolwe kuzo zonke izici.Ekugcineni, konke ukukhuliswa kuqediwe, futhi imiphumela kufanele yethulwe nge-algorithm yedatha eqinile ukuze kutholwe inani elifanele le-Ct.
Umugqa we-CT
Isinyathelo esincane sokuthuthukisa inani le-Ct izinyathelo ezingenakubalwa zabasebenzi be-R&D.Ekukhathazekeni nasekujabuleni kwemini nobusuku, njalo lapho “izintaba nemifula kungabonakali ndawo, iminyezane imnyama nezimbali zikhanya futhi kukhona omunye umuzi”, okusinika isibindi nothando lokuqhubekela phambili. futhi.


Isikhathi sokuthumela: Sep-29-2022