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ELISA cov khoom siv yog dab tsi?

2022-12-23

Cov khoom siv ELISA yog raws li cov theem ntawm cov tshuaj tiv thaiv kab mob los yog cov tshuaj tiv thaiv kab mob thiab enzyme daim ntawv lo ntawm antigen los yog tshuaj tiv thaiv. Lub antigen lossis antibody khi rau saum npoo ntawm cov khoom thauj khoom tseem khaws nws cov kev tiv thaiv kab mob, thiab cov enzyme sau npe antigen los yog cov tshuaj tiv thaiv kab mob khaws cia ob qho tib si nws cov kev tiv thaiv kab mob thiab kev ua haujlwm enzyme. Thaum lub sijhawm txiav txim siab, cov qauv kuaj hauv qab no (hauv qhov ntsuas cov tshuaj tiv thaiv kab mob lossis cov tshuaj tiv thaiv antigen) cuam tshuam nrog cov tshuaj tiv thaiv kab mob lossis cov tshuaj tiv thaiv kab mob ntawm cov khoom thauj khoom. Lub antigen-antibody complex tsim los ntawm cov khoom thauj khoom yog sib cais los ntawm lwm yam khoom hauv cov kua los ntawm kev ntxuav.

Enzyme-labeled antigens lossis cov tshuaj tiv thaiv kab mob ntxiv, uas kuj khi rau cov khoom thauj khoom los ntawm cov tshuaj tiv thaiv. Nyob rau lub sijhawm no, tus nqi ntawm cov enzyme nyob rau hauv cov khoom theem yog nyob rau hauv kev faib ua feem rau tus nqi ntawm cov khoom nyob rau hauv cov qauv. Tom qab ntxiv cov substrate ntawm cov tshuaj tiv thaiv enzyme, lub substrate yog catalyzed los ntawm enzyme los ua cov khoom xim. Tus nqi ntawm cov khoom yog ncaj qha ntsig txog tus nqi ntawm cov tshuaj ntsuam xyuas hauv cov qauv, yog li kev soj ntsuam zoo lossis ntau npaum li cas tuaj yeem ua raws li qhov tob ntawm cov xim.

Lub siab catalytic efficiency ntawm enzymes indirectly amplifies cov txiaj ntsig ntawm lub cev tiv thaiv kab mob, ua rau qhov kev ntsuam xyuas siab heev. ELISA tuaj yeem siv los txiav txim siab antigens, tab sis kuj tuaj yeem siv los txiav txim cov tshuaj tiv thaiv.

Cov hauv paus ntsiab lus ntawm ELISA cov khoom siv
Nws siv cov tshuaj tiv thaiv tshwj xeeb ntawm cov tshuaj tiv thaiv kab mob thiab cov tshuaj tiv thaiv kab mob los txuas cov khoom mus rau enzyme, thiab tom qab ntawd tsim cov tshuaj tiv thaiv xim ntawm enzyme thiab substrate rau kev txiav txim siab ntau. Cov khoom ntawm kev ntsuas tuaj yeem yog cov tshuaj tiv thaiv kab mob lossis antigen.

Muaj peb yam reagents tsim nyog hauv txoj kev txiav txim siab no:
â, Ib theem antigen lossis antibody (immune adsorbent)
Enzyme labeled antigen los yog antibody (marker)
â¢ substrate rau enzyme kev txiav txim (xim tsim tus neeg sawv cev)

Hauv kev ntsuas, cov tshuaj tiv thaiv kab mob (antibody) yog thawj zaug khi rau cov khoom thauj khoom, tab sis tseem khaws nws lub cev tiv thaiv kab mob, thiab tom qab ntawd muaj kev sib txuas (kos npe) ntawm cov tshuaj tiv thaiv kab mob (antigen) thiab enzyme ntxiv, uas tseem khaws nws qhov qub kev tiv thaiv kab mob thiab enzyme. kev ua si. Thaum lub conjugate reacts nrog cov antigen (antibody) ntawm cov khoom thauj khoom, qhov sib thooj substrate ntawm cov enzyme ntxiv. Ntawd yog, catalytic hydrolysis lossis REDOX cov tshuaj tiv thaiv thiab xim.

Qhov ntxoov ntxoo ntawm cov xim nws tsim yog proportional rau tus nqi ntawm antigen (antibody) los ntsuas. Cov khoom siv xim no tuaj yeem pom los ntawm qhov muag liab qab, kho qhov muag tsom iav, hluav taws xob microscope, kuj tuaj yeem ntsuas los ntawm spectrophotometer (enzyme label instrument). Cov txheej txheem yog yooj yim, yooj yim, ceev thiab meej.

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