I-Bovine Tuberculosis Antibody Elisa Kit

Ukucaciswa

BTB Ab Elisa

Ukusetshenziswa Okuhlosiwe: I-Bovine Tuberculosis Antibody ELISA Detection Kit ingeyokunquma isifo sofuba sezinkomo

I-antibody ku-serum yezinkomo ye-Mycobacterium bovis in vitro.

Ukuhlolwa kusetshenziselwa ukuhlola ukutheleleka kwesifo sofuba sezinkomo. I-MPB70-MPB83 i-chimera recombinant isetshenzisiwe kulokhu kuhlolwa.

Isifanekiso: serum

Izinto Ezidingekayo Kodwa Azinikezwa:

Amanzi ahlanzekile

I-Pipettor

Isifundi seMicroplate esingu-96-well (450nm)

Isikhathi

I-Centrifuge (4000rpm)

Umshini wokuwasha

Ihhavini (37 degrees)

Izinqubo Zokuhlola Nokuhumusha

ISINYATHELO 1Inqubo Yokuhlola

1.1 Ukulungiselela Isixazululo Sokugeza

Wukuhlushwa kwe-orking wi-ashing liquid:Ngaphambi kokusetshenziswa, isisombululo sokugeza kufanele sibuyiselwe ekamelweni lokushisa (20-25 degrees) futhi sinyakaziswe ngokulinganayo ukuze senze okufanayo. Bese uhlanza izikhathi ezingu-20 isisombululo sokugeza esigxiliwe ngamanzi ahlanzekile (isibonelo, isixazululo sokugeza esigxilile esingu-5ml sengezwa ku-95ml amanzi ahlanzekile), hlanganisa kahle, futhi uketshezi lokugeza oluhlanjululwe lungagcinwa ku-2-8℃izinsuku ezingu-7.

1.2 Ukulungiselela Isampula

① Igazi lesilwane lonke lathathwa, lafakwa i-centrifused ku-4000rpm imizuzu engu-10, futhi kwaqoqwa amandla amakhulu. I-Serum ingabuye ihlanganiswe ngokwemvelo ngaphandle kwe-hemolysis.

② Nciphisa isampula ye-serum 1:100 (engeza isampula ye-serum engu-2ul ku-198ul yesampula ye-diluent) bese uxuba kahle.

Qaphela: Izilawuli ezingezinhle nezihle akumele zihlanjululwe.

1.3 Ukulungiselela i-enzyme conjugate

Nciphisa i-enzyme i-conjugate 1:200 (engeza i-5ul enzyme conjugate(x100) ku-995ul yesampula ye-diluent) bese uxuba kahle.

1.4 Inqubo Yokuhlola

①Thatha ilebula ye-enzyme ye-Microplate (ngokwenani lamasampuli, ingahlakazwa futhi isetshenziswe izikhathi eziningana), engeza i-100ul ye-serum ehlanjululwe kulebula ye-enzyme ye-Microplate, usethe imithombo emibili ngayinye yamasampuli angalungile namasampula, bese wengeza i-100ul ngayinye ngezilawuli ezingezinhle nezihle ekuphenduleni kahle ngokulandelanayo. Gcoba ngobumnene isampula emthonjeni, usebenzisa ifilimu yepuleti lokuvala ukuvala ipuleti, bese ufukamela ku-37℃imizuzu engu-30.

②Vula ifilimu yepuleti lokuvala, vuthulula isixazululo epuletini kahle, engeza u-300µl woketshezi lokuwasha lokugxilisa okusebenzayo emthonjeni ngamunye, bese ulahla uketshezi emthonjeni. Phinda lezi zinyathelo ezingenhla izikhathi ezi-5, bese ekugcineni ubambe ephepheni elihlanzekile elimuncayo.

③Engeza u-100µl we-Enzyme Conjugate emthonjeni ngamunye, vala ipuleti, futhi ufukamele ku-37℃imizuzu engu-45.

④Vula ifilimu yepuleti lokuvala, vuthulula isixazululo epuletini kahle, engeza u-300µl woketshezi lokugeza okugxilile osebenzayo emthonjeni ngamunye, bese ulahla uketshezi emthonjeni. Phinda lezi zinyathelo ezingenhla izikhathi ezi-5, bese ekugcineni ubambe ephepheni elihlanzekile elimuncayo.

⑤Engeza isixazululo se-substrate esingu-100µl emthonjeni ngamunye, vala ipuleti bese ufukamela imizuzu engu-10 nge-37℃kude nokukhanya.

⑥Engeza isisombululo sokumisa esingu-50µl emthonjeni ngamunye, bese unquma umphumela phakathi nemizuzu emi-5.

ISINYATHELO 2 Ukuhunyushwa komphumela

Inani le-OD450 lomthombo ngamunye lafundwa kumfundi we-microplate we-96-well.

2.1 Ukuqinisekisa

Isilinganiso senani le-OD450nm lokulawula okungekuhle kwaba<0.5 and the average OD450nm value of positive control was ≥1, the experimental results were valid. Otherwise, the test should be repeated.

2.2 Indlela yokubala

Kusampula ngayinye, bala i-S/P ngendlela elandelayo usebenzisa isampula elilungisiwe kanye namanani okulawula:

2.3 Ukuhunyushwa Kwemiphumela

Lapho i-S/P<0.25, it is judged as negative.

Uma i-S/P Inkulu noma ilingana no-0.30, kuthathwa ngokuthi iphozithivu.

Lapho 0.25 Ngaphansi noma kulingana nokuthi S/P<0.30, it is judged as suspicious, and suggest to repeat sampling and test one week later.

Amashadi ashisayo: ikhithi yenkomo yofuba i-elisa, i-China bovine tuberculosis antibody elisa abahlinzeki, abakhiqizi, imboni