I-Bovine Tuberculosis Antibody Elisa Kit

Inkcazo

BTB Ab Elisa

Usetyenziso olucetywayo: IKithi yokuFumana iBovine Tuberculosis ELISA yeyokumisela isifo sephepha senkomo.

i-antibody kwiserum yenkomo yeMycobacterium bovis in vitro.

Uvavanyo lusetyenziselwa ukuvavanya usulelo lwesifo sephepha senkomo. I-MPB70-MPB83 i-chimera recombinant isetyenziswe kolu vavanyo.

Umzekelo: serum

Izinto ezifunekayo kodwa azibonelelwanga:

Amanzi ahlambulukileyo

Pipettor

I-96-well Microplate reader (450nm)

Isibali-xesha

I-Centrifuge (4000rpm)

Imeshini yovasa Impahla

I-oveni (amadigri angama-37)

Iinkqubo zovavanyo kunye nokutolika

INYATHELO 1Inkqubo yoVavanyo

1.1 Hlamba Ukulungiselela Isisombululo

Working concentration wulwelo oluthuthu:Ngaphambi kokusetyenziswa, isisombululo sokuhlamba kufuneka sibuyiselwe kwindawo yokushisa (i-20-25 degree) kwaye ishukunyiswe ngokulinganayo ukuze ifane. Emva koko uhlambulule amaxesha angama-20 isisombululo sokuhlamba esixutywe ngamanzi ahlambulukileyo (umzekelo, i-5ml isisombululo sokuhlamba esigxininisiweyo songezwa kwi-95ml yamanzi ahlambulukileyo), xuba kakuhle, kwaye i-liquid yokuhlamba i-diluted ingagcinwa kwi-2-8℃iintsuku ezi-7.

1.2 Ukulungiselela iSampuli

① Igazi elipheleleyo lesilwanyana lathathwa, lafakwa i-centrifuged kwi-4000rpm imizuzu eyi-10, kwaye i-supernatant yaqokelelwa. I-Serum inokudityaniswa ngokwemvelo ngaphandle kwe-hemolysis.

② Nciphisa isampuli ye-serum 1: 100 (yongeza i-2ul isampuli ye-serum kwi-198ul isampuli ye-diluent) kwaye udibanise ngokucokisekileyo.

Qaphela: Ulawulo olubi kunye nolulungileyo akufuneki lungxengwe.

1.3 ULungiselelo lwe-enzyme conjugate

Nciphisa i-enzyme i-conjugate 1: 200 (yongeza i-5ul enzyme conjugate (x100) kwi-995ul isampuli ye-diluent) kwaye udibanise ngokucokisekileyo.

1.4 Inkqubo yoVavanyo

①Thatha i-enzyme ileyibhile i-Microplate (ngokwenani leesampulu, inokuchithwa kwaye isetyenziswe ngamaxesha amaninzi), yongeza i-100ul ye-serum ehlanjululweyo kwileyibhile ye-enzyme ye-Microplate, setha amaqula amabini kwisampulu ezingalunganga nezilungileyo, kwaye ungeze i-100ul nganye yolawulo olubi kunye nolulungileyo kwi-reaction ngokulandelelana. Gcoba ngobumnene isampuli emthonjeni, usebenzisa ifilimu yokutywina ukuvala iplate, kwaye ugxininise kwi-37℃imizuzu engama-30.

②Vula ifilimu yepleyiti yokutywina, vuthulula isisombululo kwipleyiti kakuhle, yongeza i-300µl yolwelo lokuxinanisa olusebenzayo kwiqula ngalinye, emva koko ulahle ulwelo equleni. Phinda la manyathelo angasentla amaxesha ama-5, kwaye ekugqibeleni uyomise kwiphepha elicocekileyo elifunxayo.

③Yongeza i-100µl ye-Enzyme Conjugate kwiqula ngalinye, gquma ipleyiti, kwaye ufukame kwi-37℃imizuzu engama-45.

④Vula ifilimu yepleyiti yokutywina, vuthulula isisombululo kwipleyiti kakuhle, yongeza i-300µl yolwelo loxinzelelo olusebenzayo kwiqula ngalinye, emva koko ulahle ulwelo equleni. Phinda la manyathelo angasentla amaxesha ama-5, kwaye ekugqibeleni uyomise kwiphepha elicocekileyo elifunxayo.

⑤Yongeza i-100µl isisombululo se-substrate kwiqula ngalinye, vala ipleyiti kwaye ufukamele imizuzu eyi-10 nge-37℃kude nokukhanya.

⑥Yongeza 50µl isisombululo sokumisa kwiqula ngalinye, kwaye umisele isiphumo kwimizuzu emi-5.

INYATHELO 2 Isiphumo sokutolika

Ixabiso le-OD450 lequla ngalinye lafundwa kwi-96-well microplate reader.

2.1 Ukuqinisekiswa

Umndilili we-OD450nm wexabiso lolawulo olubi<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

Kwisampulu nganye, bala iS/P ngolu hlobo lulandelayo usebenzisa isampulu elungisiweyo kunye namaxabiso olawulo:

2.3 Isiphumo sokutolika

Xa iS/P<0.25, it is judged as negative.

Xa i-S/P Inkulu kuno okanye ilingana no-0.30, ithathwa njenge-positive.

Xa 0.25 Ngaphantsi okanye elingana no S/P<0.30, it is judged as suspicious, and suggest to repeat sampling and test one week later.

Iifayile ezishisayo: ibhovine tuberculosis antibody elisa kit, China bovine tuberculosis antibody elisa kit ababoneleli, abavelisi, umzi-mveliso