Qhov Tseem Ceeb Qhov txawv - Direct vs Indirect ELISA
Ib qho enzyme-txuas immunoassay (ELISA), tseem hu ua enzyme immunoassay, yog ib qho kev kuaj serological uas kuaj pom cov tshuaj tiv thaiv hauv cov ntshav. Nws yog siv los ua cov cuab yeej kuaj mob kom paub seb tus neeg mob puas tau kis tus kab mob tshwj xeeb lossis lwm yam kab mob (antigen) thiab seb lub cev puas tau tsim cov tshuaj tiv thaiv kab mob. ELISA tseem tuaj yeem txheeb xyuas cov kab mob yav dhau los thiab tam sim no. Li no, ELISA feem ntau yog siv los ua qhov kev ntsuam xyuas ua ntej los ntawm cov kws kho mob ua ntej kev tshuaj xyuas qhov tob ntawm cov kab mob. Qhov kev kuaj no tuaj yeem ua tau hauv chav kuaj los ntawm kev kuaj ntshav los ntawm tus neeg mob. Muaj ob hom kev xeem ELISA: ncaj qha ELISA thiab tsis ncaj ELISA. Qhov sib txawv tseem ceeb ntawm kev ncaj qha thiab tsis ncaj ELISA yog qhov ncaj qha ELISA yog qhov nkag siab ntau dua thiab yuav tsum tau ntxiv cov tshuaj tiv thaiv kab mob thib ob thaum ncaj qha ELISA tsis tshua nkag siab thiab tsuas yog siv cov tshuaj tiv thaiv thawj zaug xwb.
Direct ELISA yog dab tsi?
ELISA yog ib qho kev kuaj mob uas tau ua los txheeb xyuas qhov muaj cov tshuaj tiv thaiv tshwj xeeb lossis cov tshuaj tiv thaiv kab mob hauv cov ntshav. Nws yog ua raws li daim ntawv ntsuam xyuas. Nws siv cov tshuaj tiv thaiv kab mob txuas nrog cov tshuaj enzymes yooj yim. Antigens muaj nyob rau hauv cov qauv ntshav khi nrog cov tshuaj tiv thaiv tshwj xeeb txuas nrog enzymes. Nyob rau hauv cov kauj ruam kawg, ib tug tshwj xeeb substrate yog ntxiv rau react nrog lub enzyme. Lub enzyme hloov lub substrate mus rau hauv ib qho xim los yog teeb liab tsim khoom. Cov xim hloov hauv cov tshuaj substrate qhia pom muaj cov tshuaj tiv thaiv tshwj xeeb hauv cov qauv ntshav. Direct ELISA test siv tsuas yog thawj cov tshuaj tiv thaiv kab mob txuas nrog cov enzyme. Thaum khi rau cov antigen, nws hloov xim sai sai, qhia tias muaj tus kab mob hauv cov ntshav. Txawm li cas los xij, qhov kev siv ntawm cov teeb liab tsis muaj zog hauv ELISA ncaj qha vim cov epitopes raug txwv rau kev khi ntawm cov antigens. Yog li ntawd, ncaj qha ELISA tsis tshua nkag siab piv rau qhov tsis ncaj ELISA.
Daim duab 01: Direct ELISA Test
ELISA Indirect yog dab tsi?
ELISA tuaj yeem ua tau siv ob hom tshuaj tiv thaiv uas yog; thawj antibody thiab Secondary antibody. Indirect ELISA cov cuab yeej siv ob hom tshuaj tiv thaiv kab mob kom nthuav dav cov cim rau kev tshawb pom zoo dua. Indirect ELISA cov txheej txheem yog ua raws li hauv qab no.
- Plates yog incubated nrog antigens thiab ntxuav los thaiv nonspecific binding.
- Tom qab ntawd cov tshuaj tiv thaiv thawj zaug ntxiv thiab ntxuav.
- Enzyme-linked Secondary antibody ntxiv thiab ntxuav.
- Ib lub substrate tau ntxiv thiab tso cai rau hnov mob nrog enzymes.
- Signals raug kuaj pom, thiab muaj lossis tsis muaj cov tshuaj tiv thaiv tshwj xeeb hauv cov qauv raug txheeb xyuas.
Hauv kev sim ELISA tsis ncaj, ntau cov tshuaj tiv thaiv kab mob thib ob tuaj yeem khi rau ib qho tshuaj tiv thaiv thawj zaug. Cov tshuaj tiv thaiv thib ob yog txuas nrog cov enzymes yooj yim soj ntsuam. Yog li ntawd, ib qho kev khi tuaj yeem ua rau lub teeb liab muaj zog vim muaj ntau tshaj ib qho kev sib cuam tshuam. Li no, tsis ncaj ELISA yog qhov nkag siab ntau dua li ncaj qha ELISA. Txawm li cas los xij, tsis ncaj ELISA tuaj yeem ua cov cim tsis tshwj xeeb vim muaj kev cuam tshuam ntawm cov tshuaj tiv thaiv thib ob.
Daim duab 02: Indirect ELISA Test
Qhov txawv ntawm Direct thiab Indirect ELISA yog dab tsi?
Direct vs Indirect ELISA |
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| Direct ELISA yog qhov tsis tshua muaj txiaj ntsig piv rau qhov tsis ncaj ELISA. | Indirect ELISA rhiab dua. |
| Direct ELISA xeem yog txheej txheem ceev. | Indirect ELISA yog lub sijhawm siv. |
| siv tshuaj tiv thaiv | |
| Tsuas yog ib hom tshuaj tiv thaiv kab mob (Primary antibodies) raug siv ncaj qha ELISA. | Cov tshuaj tiv thaiv thawj thiab theem nrab yog siv rau kev tsis ncaj ncees ELISA. |
| Txuas nrog Enzymes | |
| Cov tshuaj tiv thaiv thawj zaug yog txuas nrog enzymes. | Cov tshuaj tiv thaiv thib ob yog txuas nrog enzymes. |
| Cross-Reactivity of Second Antibodies | |
| Direct ELISA tshem tawm qhov sib txawv ntawm cov tshuaj tiv thaiv thib ob. | Indirect ELISA cuam tshuam nrog kev sib cuam tshuam ntawm cov tshuaj tiv thaiv thib ob. |
| Signals | |
| Signals tsis muaj zog piv rau indirect ELISA. | Signals yog amplified indirect ELISA. Li no, nws yog ib qho yooj yim mus ntes. |
Summary – Direct vs Indirect ELISA
ELISA yog cov txheej txheem biochemical siv feem ntau hauv kev tiv thaiv kab mob txhawm rau kuaj pom muaj cov tshuaj tiv thaiv kab mob lossis cov tshuaj tiv thaiv kab mob hauv cov ntshav ntawm tus neeg mob. Nws tuaj yeem ua tau los ntawm ob txoj haujlwm hu ua ELISA ncaj qha lossis ncaj qha. Direct ELISA test siv tsuas yog thawj cov tshuaj tiv thaiv kab mob txhawm rau txheeb xyuas cov antigen thaum tsis ncaj ELISA siv cov tshuaj tiv thaiv thawj thiab theem nrab. Hauv ELISA ncaj qha, thawj cov tshuaj tiv thaiv kab mob tau sau npe thaum tsis ncaj ncees ELISA cov tshuaj tiv thaiv kab mob thib ob tau sau npe. Qhov no yog qhov txawv ntawm qhov ncaj thiab tsis ncaj ELISA.
