Yamamolekhula Diagnostics: real-time-PCR

Okwamanje, ukuhlola yamangqamuzana indlela yakhelwe polymerase chain reaction (i-PCR), ebelikude umfutho sokuthuthukisa inani elikhulu izindlela ukuxilongwa izifo, esekelwe ukuzimisela zofuzo. Sokufeza real-time-PCR usebenzisa isakhi sofuzo isinqamu (ikakhulukazi i-nucleic acid) ye-ejenti causative, okuyinto ayitholakali abanye abantu.

Umgomo we-PCR iyona ukubuyabuyelela lwegciwane DNA isinqamu izigidi ezithile zesikhathi DNA polymerase e uphiko oluyisipesheli, ukubizwa ngokuthi cycler ezishisayo, esihlinzeka ukuphindaphinda eyindilinga umbuso lokushisa for uhlelo unikezwa.

Ukuxilonga i-PCR ivumela ukuphinda uchungechunge amplifications, ukugcina umsebenzi-enzyme. Lapho okhulisa senzeka ngokushelela, komunye umjikelezo inani amakhophi ingxenye ethize yalokho DNA kabili lwegciwane, ngemva amathathu nowe - a neshumi. Yingakho i-PCR isetshenziswa uphethwe ukunquma Amagciwane sincane ngokwanele, ukuhlonza zazo akunakwenzeka ngaphandle kwalo.

Kufanele kuqashelwe ukuthi i-RNA hhayi kungaba kuyisibonelo PCR ngesikhathi sangempela. Ngakho-ke, ukuze sithole RNA viral phambi okhulisa ukuze DNA kuhlobene nezinga isinqamu ohambelana ke. Ukusabela polymerase chain ivumela ukukhiqiza i-DNA ne-RNA nesilinganiso lwegciwane. Ulwazi ngaleyo ndlela etholwe isetshenziselwa ukuqapha ukusebenza yokwelapha esetshenziswa, kanye nokuhlola onalesi sifo emitholampilo.

Ngaphezu kwalokho, real-time-PCR akudingi izenzo ezengeziwe mayelana nencazelo ye-DNA target noma amplicons, okuyizindawo yisithiyo uphethwe yini futhi kungaholela emiphumeleni zamanga. Le ndlela kwenza kube nokwenzeka ukuba angasebenzisi isinyathelo electrophoresis, okuholela imikhiqizo zokunciphisa ingcuphe yokutheleleka ukuthi bayavuma polymerase chain. Futhi, ngenxa ukwehla kwenani manipulations ne-ejenti, kunciphisa isikhathi inhlolovo, yenza lula inqubo, kunciphisa amathuba amaphutha. PCR nako ikuvumela ukuba bacacise inani kokuqala amakhophi DNA lwegciwane e zesivivinyo sasemtholampilo ukusiza ukukhomba izinkathi exacerbations futhi ukuthatha izinyathelo ngesikhathi esifanele ukwelapha isiguli.

Namuhla, kukhona inani elikhulu izindlela yokukhulisa nenkanuko edlulele ngesikhathi sangempela. Ezisetshenziswa kakhulu amasu ahlukahlukene Ukusebenzisa amplicon kuhlobene nezinga iphenya fluorogenic, namandla emisebe ye-zazo iyahlukahluka kuye ngokuthi ekuqongeleleni amplicon futhi iqoshwe ngesikhathi sangempela esebenzisa idivayisi elikhethekile okuthiwa i thermocycler nge-module optical. Linikeza ezingeni lelisetulu ukuzwela yokuhlaziya. Ngesikhathi umkhiqizo ofanayo PCR kungatholakala, kanye inqwaba by-imikhiqizo, ngesikhathi lapho usebenzisa electrophoresis ke kwezinye izimo waphawula.

Lapho usebenzisa i-PCR amasampula omuhle nezilawuli ezingezinhle ngesikhathi sangempela. Ngakho, amasampula emihle nemibi esifundweni ihlaziye amasampula ahlukene, isampula ukulawula lapho kwetfulwa isampula ngalinye okunjalo futhi sihamba zonke izinyathelo ukuhlaziywa.

Namuhla, kukhona inani elikhulu kumadivayisi sokufeza ukusabela polymerase chain. Lezi zihlanganisa Bio-Rad, osetshenziswayo Biosystems, Cepheid futhi Roche. Ngamunye wabo ngebuhle nebubi

Kufanele kuqashelwe ukuthi i-PCR - indlela uphethwe elisetshenziswa emikhulu yesayensi, ucwaningo kanye izikhungo zokuxilonga emhlabeni wonke, ngoba kunika ithuba lokufeza incazelo ngokushesha imiphumela.