Ko taw thiab tus kab mob ko taw-o tshuaj tiv thaiv Elisa Cov Khoom Siv

Kev qhia tshwj xeeb

Fmdv-o ab elisa

Siv: Lub bovine ko taw thiab qhov ncauj tus kab mob-o

Tshuaj tiv thaiv cov tshuaj tiv thaiv ELISA nrhiav tau rau kev txiav txim siab ntawm ko taw thiab qhov ncauj tiv thaiv hauv Bibine cov ntshav dej hauv vitro. Qhov kev xeem no yog siv los ntsuas tus kab mob ko taw thiab qhov chaw ntawm cov tshuaj tiv thaiv kab mob o antibodity kev tiv thaiv kev ua liaj ua teb.

Ib yam: ntshav qab zib

Cov ntaub ntawv yuav tsum tau tab sis tsis muab rau:

Lim Dej

Lub tso dej

{{0 {}} Zoo Microplate nyeem ntawv (450nm)

Timer

Centrifuge (4000rpm)

Tshuab ntxhua khaub ncaws

CUB (37 DEGREE)

Cov Txheej Txheem Kuaj & Kev Txhais Lus

Kauj ruam 1 Kuaj cov txheej txheem

1.1 Npaj Ua Tau

Working concentration washing ua kua:Ua ntej siv, kev ua tiav kev ntxuav yuav tsum tau rov qab mus txog chav sov (20-25 degree) thiab shaken tusyees los ua kom nws tsis zoo. Tom qab ntawd dilute 20 lub sij hawm lub rauv ntxuav nrog cov dej ntxuav (piv txwv, 5ml zoo, thiab siv cov kua ntxuav tawm tuaj yeem khaws cia ntawm {}} degree rau 7 hnub.

1.2 Kev Npaj Qauv Qauv

① Tag nrho cov tsiaj cov ntshav tau tshwm sim, centrifuged ntawm 4000RPM rau 10 feeb, thiab supernatant tau sau. Ntshav dej kuj tseem tuaj yeem ua rau muaj kev cuam tshuam ib txwm tsis muaj hemolysis.

② Dilute Dilute Cov Qauv Sampution 1:25 (Ntxiv 5ul qauv tshuaj rau 120ul coj mus ua piv txwv) thiab sib tov kom huv si.

Nco tseg: tsis zoo thiab kev tswj hwm zoo kuj yuav tsum muaj dilute.

1.3 enzyme conjugate npaj

Dilute cov enzyme conjugate 1:90 (ntxiv 5ul enzyme convugate (x100) coj mus rau 445ul cojilsilent) thiab sib tov kom huv si.

1.4 Cov Txheej Txheem Ntsuas

①Take Tus Enzyme sau microplate (raws li tus naj npawb ntawm cov qauv, muab ob peb zaug rau cov qauv tsis zoo thiab muaj txiaj ntsig zoo rau qhov tsis zoo thiab kev tswj hwm zoo rau cov tshuaj tiv thaiv tau raws li. Maj mam muab cov qauv hauv lub qhov dej, siv cov phaj sealing los npog lub phaj, thiab ncu ntawm 37 degree rau 30 feeb.

Oopen cov ntawv sib khi ua yeeb yaj kiab, co tawm ntawm kev daws hauv lub phaj zoo, ntxiv 3004L ntawm kev ua haujlwm siab ntxuav kua rau txhua qhov, thiab tom qab ntawd pov tseg cov kua hauv lub qhov dej. Rov ua cov kauj ruam saum toj no 5 zaug, thiab thaum kawg pat qhuav ntawm daim ntawv huv si.

③add 100μl ntawm enzyme conjugate rau txhua qhov zoo, npog phaj, thiab ncos ntawm 37 degree rau 45 feeb.

Oopen cov ntawv sib khi ua yeeb yaj kiab, co tawm ntawm kev daws hauv lub phaj zoo, ntxiv 3004L ntawm kev ua haujlwm siab ntxuav kua rau txhua qhov, thiab tom qab ntawd pov tseg cov kua hauv lub qhov dej. Rov ua cov kauj ruam saum toj no 5 zaug, thiab thaum kawg pat qhuav ntawm daim ntawv huv si.

⑤add 100μL Substrate daws rau txhua lub phaj thiab ncos rau 10 feeb ntawm 37 degree deb ntawm lub teeb.

⑥add 50μl nres kev daws rau txhua qhov zoo, thiab txiav txim siab qhov tshwm sim tsis pub dhau 5 feeb.

QEEM 2 TIM Txhais Lus

OD450 tus nqi ntawm txhua qhov dej tau nyeem nyob rau ntawm {1}}} microplate nyeem ntawv.

2.1 kev siv tau

Qhov nruab nrab Od450nm tus nqi ntawm kev tswj hwm tsis zoo yog<0.15 and the average OD450nm value of positive control /the average OD450nm value of negative control was ≥3, the experimental results were valid. Otherwise, the test should be repeated.

2.2 Kev Xam Xyuas Xam

Rau txhua tus qauv, laij tus s / p raws li hauv qab no siv cov qauv kho thiab tswj qhov tseem ceeb:

2.3 Qhov txhais Kev Txhais Lus

Thaum s / p<0.15, it is judged as negative.

Thaum s / p ntau dua los yog sib npaug rau 0. 25, nws raug txiav txim raws li qhov zoo.

Thaum {0. 15 tsawg dua lossis sib npaug rau S / P<0.25, it is judged as suspicious, and suggest to repeat sampling and test one week later.

Cim npe nrov: Ko taw thiab tus kab mob ko taw-o tshuaj tiv thaiv Elisa Cov Khoom Siv, Tuam Tshoj thiab qhov ncauj kab mob-O Antibody Elisa cov khoom siv, chaw tsim tshuaj paus, Hoobkas