Qhov sib txawv tseem ceeb ntawm Elisa thiab Elispot yog Elisa yog ib qho enzyme-txuas immunosorbent assay uas txiav txim siab tag nrho cov concentration ntawm secreted signaling proteins, thaum Elispot yog ib tug enzyme-txuas immunosorbent assay uas kuaj ib tug neeg cytokine-secreting hlwb.
Elisa thiab Elispot yog ob qhov kev tshuaj ntsuam enzyme-txuas immunosorbent dav siv hauv tshuaj, nroj tsuag pathology, biotechnology thiab kev lag luam tshuaj sib txawv. Nyob rau hauv cov tswv yim no, ib qho tshuaj ntsuam xyuas protein yog kuaj los ntawm cov tshuaj tiv thaiv kab mob tiv thaiv nws. Elisa yog qhov nrov tshaj plaws enzyme-txuas immunosorbent assay siv hauv chav kuaj. Ntawm qhov tod tes, Elispot yog ib qho enzyme-linked immunosorbent assay uas tsis yog siv los ntawm Elisa tab sis siv ntxiv rau Elisa hauv chav kuaj.
Elisa Assay yog dab tsi?
Elisa yog ib qho enzyme-txuas immunosorbent assay uas txiav txim siab tag nrho cov concentration ntawm secreted signaling proteins. Nws yog ib qho kev tshuaj ntsuam xyuas biochemistry uas tau siv dav dav uas tau piav qhia thawj zaug los ntawm Engvall thiab Perlmann hauv xyoo 1971. Cov txheej txheem no siv cov txheej txheem ntawm cov enzyme immunoassay txhawm rau kuaj pom muaj cov protein tshwj xeeb hauv cov qauv ua kua siv cov tshuaj tiv thaiv kab mob tiv thaiv cov protein tshwj xeeb no. Qhov kev ntsuam xyuas no yog ib qho cuab yeej siv tshuaj ntsuam xyuas hauv cov tshuaj, cog pathology thiab biotechnology.
Mechanism of ELISA Test
Nyob rau hauv feem ntau ntawm ELISA xeem, ib qho tsis paub tus nqi ntawm cov tshuaj tiv thaiv yog immobilized mus rau cov khoom txhawb nqa (microlitre phaj) thaum pib. Tom qab ntawd qhov sib txuam thawj cov tshuaj tiv thaiv yog siv rau ntawm qhov chaw txhawb nqa khoom. Cov tshuaj tiv thaiv no tsim ib qho kev sib txuam nrog antigen (antigen-antibody complex). Lub antigen khi thawj cov tshuaj tiv thaiv tuaj yeem kuaj pom los ntawm cov tshuaj tiv thaiv kab mob thib ob uas txuas rau ib qho enzyme los ntawm bioconjugation. Hauv cov kauj ruam kawg ntawm kev sim Elisa, ib yam khoom uas muaj cov substrate ntawm cov enzyme ntxiv. Yog tias muaj kev sib khi zoo ntawm cov tshuaj tiv thaiv thawj zaug rau cov antigen, cov xim hloov tau raug tsim los ntawm substrate. Thaum kawg, qhov no suav cov antigen hauv cov qauv.
Daim duab 01: Ntau hom Elisa
Tsis tas li ntawd, muaj ntau txoj hauv kev rau Elisa, xws li ncaj qha Elisa thiab ncaj qha Elisa. Tam sim no, ntau yam kev ntsuam xyuas Elisa yog siv los kuaj tib neeg cov kab mob xws li Mycobacterium tuberculosis, rotavirus, kab mob siab B, kab mob siab C, enterotoxin ua E.coli, HIV, thiab SARS-CoV-2.
Elispot Assay yog dab tsi?
Elispot yog ib qho enzyme-txuas immunosorbent assay uas kuaj pom tus kheej cytokine-secreting hlwb. Cecil Czerkinsky thawj zaug piav ELISpot hauv xyoo 1983. Nws yog ib hom kev ntsuam xyuas uas tsom mus rau qhov ntsuas qhov ntau zaus ntawm cytokine secretion rau ib leeg. Nws tseem raug cais raws li cov txheej txheem uas siv cov tshuaj tiv thaiv kab mob los kuaj xyuas cov tshuaj lom neeg lossis tshuaj lom neeg cov protein. Fluorospot assay yog ib qho kev hloov pauv ntawm Elispot assay, uas siv cov fluorescence txhawm rau txheeb xyuas ntau yam analytes (ntau cov protein ntau zais cia).
Mechanism ntawm Elispot Test
Nyob rau hauv cov txheej txheem no, cytokine tshwj xeeb monoclonal antibodies raug muab ntxiv rau hauv qhov dej ntawm microlitre daim hlau pib. Tom qab ntawd cov hlwb xav tau raug soj ntsuam ntxiv rau hauv qhov dej. Tom qab ntawd, cov cell yog incubated. Thaum lub sij hawm incubation ntawm tes, lub hlwb raug tso cai rau hnov mob rau tej yam stimuli thiab secrete cytokine. Cov cytokines uas tau muab zais los ntawm cov hlwb incubated yuav txuas mus rau cov tshuaj tiv thaiv txij li lub hlwb raug ncig los ntawm cytokine tshwj xeeb monoclonal antibodies.
Daim duab 02: Elispot Assay
txhawm rau txheeb xyuas cov cytokine tshwj xeeb cov tshuaj tiv thaiv kab mob monoclonal, biotinylated tshwj xeeb kuaj cov tshuaj tiv thaiv raug ntxiv rau hauv qhov dej. Biotinylated tshwj xeeb kuaj cov tshuaj tiv thaiv ntes cov cytokine tshwj xeeb monoclonal antibodies hauv cov qhov dej. Ntxiv mus, streptavidin-enzyme conjugates tau ntxiv rau hauv qhov dej txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txheeb xyuas cov tshuaj tiv thaiv. Hauv cov kauj ruam kawg, cov substrates tshwj xeeb tau ntxiv rau hauv qhov dej. Cov tshuaj tiv thaiv ntawm cov enzyme conjugates thiab substrates tsim cov chaw sib txawv hauv cov qhov dej. Cov chaw sib txawv no tau nyeem ntawm tus nyeem ntawv ELISpot. Thaum kawg, qhov no yuav suav ntxiv qhov zaus ntawm cytokine secretion.
Ib yam zoo sib xws ntawm Elisa thiab Elispot?
- Lawv yog ob qho tib si enzyme-txuas immunosorbent assays.
- Ob qhov kev soj ntsuam kuaj pom cov protein analytes.
- Cov kev ntsuam xyuas no siv cov tshuaj tiv thaiv tshwj xeeb txhawm rau txheeb xyuas cov protein analytes.
- Ob qhov kev ntsuam xyuas siv enzymes thiab substrates.
- Lawv siv los ua cov cuab yeej kuaj mob hauv chaw kuaj mob.
Qhov txawv ntawm Elisa thiab Elispot yog dab tsi?
Elisa yog ib qho enzyme-linked immunosorbent assay uas txiav txim siab tag nrho cov concentration ntawm secreted signaling proteins. Ntawm qhov tod tes, Elispot yog ib qho enzyme-txuas immunosorbent assay uas ntsuas tus kheej cytokine secreting hlwb. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm Elisa thiab Elispot. Ntxiv mus, hauv kev sim Elisa, cov tshuaj tiv thaiv yog thawj zaug immobilized mus rau microlitre qhov dej. Hauv qhov sib piv, hauv kev sim Elispot, cov tshuaj tiv thaiv yog thawj zaug immobilized mus rau microlitre qhov dej. Yog li, qhov no yog lwm qhov sib txawv ntawm Elisa thiab Elispot.
Cov duab hauv qab no nthuav tawm cov ncauj lus kom ntxaws ntawm qhov sib txawv ntawm Elisa thiab Elispot hauv daim ntawv teev npe.
Summary – Elisa vs Elispot
Elisa thiab Elispot yog ob qhov kev tshuaj ntsuam enzyme-txuas immunosorbent dav siv hauv tshuaj, nroj tsuag pathology, biotechnology, thiab kev lag luam tshuaj sib txawv. Elisa kuaj pom muaj cov ligand (protein) hauv cov qauv ua kua siv cov tshuaj tiv thaiv kab mob tiv thaiv cov ligand los ntsuas. Elispot assay yog ib txoj hauv kev siv dav los saib xyuas cov tshuaj tiv thaiv kab mob. Nws tshawb pom tus kheej cytokine secreting hlwb siv cov tshuaj tiv thaiv. Nws yog ob qho tib si immunoassay thiab bioassay txheej txheem. Yog li, qhov no yog cov ntsiab lus ntawm qhov sib txawv ntawm Elisa thiab Elispot.
