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Yeyiphi imisebenzi yekhithi ye-ELISA?
2022-12-23
Ikhithi ye-ELISA isekwe kwisigaba esiluqilima se-antigen okanye i-antibody kunye nokuleyibhelishwa kwe-enzyme ye-antigen okanye i-antibody. I-antigen okanye i-antibody ebotshelelwe kumphezulu wesithwali esiqinileyo isawugcina umsebenzi wayo wokugonyela, kwaye i-enzyme ebizwa ngokuba yi-antigen okanye i-antibody igcina zombini umsebenzi wayo wokugonyela kunye nomsebenzi we-enzyme. Ngexesha lokumiselwa, i-specimen ephantsi kovavanyo (apho i-antibody okanye i-antigen ilinganiswa) isabela kunye ne-antigen okanye i-antibody kumphezulu womthwali oqinileyo. I-antigen-antibody complex eyenziwe kwi-carrier eqinile yahlulwe kwezinye izinto kulwelo ngokuhlamba.
Ii-antigens ezibhalwe nge-Enzyme okanye ii-antibodies zongezwa, nazo zibophelela kumthwali oqinileyo ngokusabela. Ngeli xesha, isixa se-enzyme kwisigaba esiqinileyo silingana nesixa sezinto kwi-specimen. Emva kokongeza i-substrate ye-enzyme reaction, i-substrate i-catalyzed yi-enzyme ukuba ibe yimveliso enemibala. Isixa semveliso sihambelana ngokuthe ngqo kwisixa sezinto ezivavanyiweyo kwi-specimen, ngoko ke uhlalutyo lwekhwalithi okanye ubungakanani lunokwenziwa ngokobunzulu bombala.
Ukusebenza okuphezulu kwe-catalytic ye-enzymes ngokungathanga ngqo kukhulisa iziphumo zokuphendula komzimba, okwenza uvavanyo lube buthathaka kakhulu. I-ELISA inokusetyenziselwa ukumisela ii-antigens, kodwa ingasetyenziselwa ukumisela amajoni omzimba.
Imigaqo esisiseko yekhithi ye-ELISA
Isebenzisa impendulo ethile ye-antigen kunye ne-antibody ukudibanisa into kwi-enzyme, kwaye emva koko ivelise ukusabela kombala phakathi kwe-enzyme kunye ne-substrate yokuzimisela kobungakanani. Into yokulinganisa ingaba yi-antibody okanye i-antigen.
Kukho ii-reagents ezintathu eziyimfuneko kule ndlela yokuzimisela:
â I-antigen yesigaba esiqinileyo okanye isilwa-mzimba (i-adsorbent yamajoni omzimba)
â¡ I-enzyme ebhalwe i-antigen okanye i-antibody (umakisha)
⢠i-substrate yesenzo se-enzyme (i-arhente yophuhliso lombala)
Kwimilinganiselo, i-antigen (i-antibody) iqala ngokubophelela kwi-carrier eqinile, kodwa igcina umsebenzi wayo wokuzivikela, kwaye emva koko i-conjugate (umakishi) ye-antibody (i-antigen) kunye ne-enzyme yongezwa, esayigcina umsebenzi wayo wokuqala we-immune kunye ne-enzyme. umsebenzi. Xa i-conjugate isabela kunye ne-antigen (i-antibody) kwi-carrier carrier, i-substrate ehambelanayo ye-enzyme yongezwa. Oko kukuthi, i-catalytic hydrolysis okanye i-REDOX yokusabela kunye nombala.
Ii-antigens ezibhalwe nge-Enzyme okanye ii-antibodies zongezwa, nazo zibophelela kumthwali oqinileyo ngokusabela. Ngeli xesha, isixa se-enzyme kwisigaba esiqinileyo silingana nesixa sezinto kwi-specimen. Emva kokongeza i-substrate ye-enzyme reaction, i-substrate i-catalyzed yi-enzyme ukuba ibe yimveliso enemibala. Isixa semveliso sihambelana ngokuthe ngqo kwisixa sezinto ezivavanyiweyo kwi-specimen, ngoko ke uhlalutyo lwekhwalithi okanye ubungakanani lunokwenziwa ngokobunzulu bombala.
Ukusebenza okuphezulu kwe-catalytic ye-enzymes ngokungathanga ngqo kukhulisa iziphumo zokuphendula komzimba, okwenza uvavanyo lube buthathaka kakhulu. I-ELISA inokusetyenziselwa ukumisela ii-antigens, kodwa ingasetyenziselwa ukumisela amajoni omzimba.
Imigaqo esisiseko yekhithi ye-ELISA
Isebenzisa impendulo ethile ye-antigen kunye ne-antibody ukudibanisa into kwi-enzyme, kwaye emva koko ivelise ukusabela kombala phakathi kwe-enzyme kunye ne-substrate yokuzimisela kobungakanani. Into yokulinganisa ingaba yi-antibody okanye i-antigen.
Kukho ii-reagents ezintathu eziyimfuneko kule ndlela yokuzimisela:
â I-antigen yesigaba esiqinileyo okanye isilwa-mzimba (i-adsorbent yamajoni omzimba)
â¡ I-enzyme ebhalwe i-antigen okanye i-antibody (umakisha)
⢠i-substrate yesenzo se-enzyme (i-arhente yophuhliso lombala)
Kwimilinganiselo, i-antigen (i-antibody) iqala ngokubophelela kwi-carrier eqinile, kodwa igcina umsebenzi wayo wokuzivikela, kwaye emva koko i-conjugate (umakishi) ye-antibody (i-antigen) kunye ne-enzyme yongezwa, esayigcina umsebenzi wayo wokuqala we-immune kunye ne-enzyme. umsebenzi. Xa i-conjugate isabela kunye ne-antigen (i-antibody) kwi-carrier carrier, i-substrate ehambelanayo ye-enzyme yongezwa. Oko kukuthi, i-catalytic hydrolysis okanye i-REDOX yokusabela kunye nombala.
Umthunzi wombala owuvelisayo ulingana nobungakanani be-antigen (i-antibody) ekufuneka ilinganiswe. Le mveliso enemibala inokubonwa ngeso lenyama, i-microscope ebonakalayo, i-electron microscope, inokulinganiswa nge-spectrophotometer (isixhobo seleyibhile ye-enzyme). Indlela ilula, ilungele, ikhawuleza kwaye ithile.
Ngaphambili:Intshayelelo ye Elisa Plate
