Classic Swine Fever Virus Antibody Elisa Kit

Tsanangudzo

 

 

Kuedza Maitiro & Dudziro

 

STEP 1 Test Procedure

1.1 Wash Solution Kugadzirira

Working concentration washing liquid:Usati washandiswa, mushonga wekugeza unofanirwa kudzorerwa kumhepo yekushisa (20-25 degree) uye inozununguswa zvakafanana kuti iite yunifomu. Wobva wawedzera ka20 iyo yakaomeswa yekugezesa mhinduro nemvura yakacheneswa (semuenzaniso, 5ml yakaomeswa yekugezesa mhinduro inowedzerwa kune 95ml yakanatswa mvura), sanganisa zvakanaka, uye yakanyungudutswa yekuwachisa mvura inogona kuchengetwa pa2-8 dhigirii kwemazuva manomwe.

1.2 Muenzaniso Kugadzirira

① Ropa rose remhuka rakatorwa, rakaiswa centrifuged pa4000rpm kwemaminetsi gumi, uye supernatant yakaunganidzwa. Serum inogonawo kubatanidzwa zvakasununguka pasina hemolysis.

② Diraisa iyo serum kuti iedzwe kagumi muserum dilution plate (semuenzaniso: wedzera 15ul yeserum kuti iongororwe kune 135ul sampuli diluent).

Cherechedzo: Kudzora kwakashata uye kwakanaka hakufanirwe kuderedzwa.

1.3 Test Procedure

①Tora iyo enzyme label Microplate (maererano nenhamba yemasampuli, inogona kuparadzaniswa uye kushandiswa kakawanda), wedzera 100ul ye serum yakanyungudutswa kune iyo enzyme label Microplate, isa matsime maviri ega ega ekudzora kwakashata uye kutonga kwakanaka, uye wedzera 100ul kune mhinduro zvakanaka zvichiteerana. Zvinyoro zunza muenzaniso mugomba, uchishandisa firimu rekuvhara ndiro kuvhara ndiro, uye incubate pa 37℃kwemaminitsi makumi matatu.

②Vhura firimu rekuvharisa ndiro, zunza mhinduro mundiro zvakanaka, wedzera 300µl yemvura yekugeza yekushanda patsime rega rega, uye wobva warasa mvura mutsime. Dzokorora matanho ari pamusoro apa kashanu, uye pakupedzisira pat yakaoma papepa rakachena rinopinza.

③Wedzera 100µl yeEnzyme Conjugate kutsime rega rega, vhara ndiro, uye incubate pa37℃kwemaminitsi makumi matatu.

④Vhura firimu replate rekuvhara, zunza mhinduro mundiro zvakanaka, wedzera 300µl yekushanda yekugeza yemvura patsime rega rega, uye wobva warasa mvura mutsime. Dzokorora matanho ari pamusoro apa kashanu, uye pakupedzisira pat yakaoma papepa rakachena rinopinza.

⑤Wedzera 100µl substrate mhinduro kutsime rega rega, vhara ndiro uye incubate kwemaminetsi gumi pa 37℃kure nechiedza.

⑥Wedzera 50µl yekumisa mhinduro kutsime rega rega, uye tarisa mhedzisiro mukati memaminetsi mashanu.

 

STEP 2 Result Dudziro

Iyo OD450 kukosha kwetsime rega rega yakaverengwa pa96-tsime microplate muverengi.

2.1 Kusimbiswa

Avhareji yeOD450nm kukosha kwekutonga kwakashata yaive <0.2 uye avhareji OD450nm kukosha kwekutonga kwakanaka yaive Yakakura kupfuura kana kuenzana ne0.7, mhedzisiro yekuedza yaive inoshanda. Kana zvisina kudaro, muedzo unofanirwa kudzokororwa.

2.2 Maitiro ekuverenga

Kune yega yega sampuli, verenga iyo S/P nenzira inotevera uchishandisa yakagadziridzwa sampu uye kutonga maitiro:

2.3 Dudziro yemhedzisiro

Kana S/P <0.2, inotongwa seisina kunaka.

Kana S/P> 0.3, inotongwa seyakanaka.

Kana 0.2 Zvishoma pane kana kuenzana neS/P Zvishoma pane kana kuenzana na 0.3, zvinoonekwa sezviri kufungira, uye zvinokurudzira kudzokorora sampling uye kuyedza vhiki imwe gare gare.

 

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