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Iinkqubo zovavanyo kunye nokutolika

 

INYATHELO 1 Inkqubo 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 1:50 (yongeza i-3ul isampuli ye-serum kwi-147ul isampuli ye-diluent) kwaye udibanise ngokucokisekileyo.

Qaphela: Ulawulo olubi kunye nolulungileyo lukwafuna ukuhlanjululwa.

1.3 ULungiselelo lwe-enzyme conjugate

Nciphisa i-enzyme conjugate 1: 250 (yongeza i-2ul enzyme conjugate (x100) kwi-498ul isampuli ye-diluent) kwaye udibanise ngokucokisekileyo.

1.4 Inkqubo yoVavanyo

①Thatha i-enzyme ileyibhile i-Microplate (ngokwenani leesampulu, ingachithwa 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 kwimpendulo 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 lokuxinanisa 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<0.1 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 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-0.35, ithathwa njengelungileyo.

Xa 0.25 Ngaphantsi okanye elingana no S/P<0.35, it is judged as suspicious, and suggest to repeat sampling and test one week later.

 

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