Classic swine fever virus antibody elisa kit .

Okunnyonnyola .

 

 

Enkola z'okugezesa & okutaputa .

 

Omutendera 1 Enkola y’okugezesa .

1.1 Okuteekateeka eddagala ery’okunaaba .

Working okukuŋŋaanyizibwa . wAshing Amazzi:Nga tonnaba kukozesa, eky’okunaaba kisaana okuzzibwa ku bbugumu erya bulijjo (20-25℃) era ne kikankana kyenkanyi okukifuula ekifaanagana. Oluvannyuma kendeeza ku mirundi 20 egy’okunaaba okusengekeddwa n’amazzi agalongooseddwa (okugeza, 5ml concentrated washing solution eyongerwa mu 95ml amazzi agalongooseddwa), tabula bulungi, era amazzi g’okunaaba agafuukuuse gasobola okuterekebwa ku 2-8℃okumala ennaku 7.

1.2 Okuteekateeka sampuli .

1 Omusaayi gw’ebisolo gwonna gwatwalibwa, nga gufuumuulwa ku 4000rpm okumala eddakiika 10, era supernatant yakuŋŋaanyizibwa. Serum era esobola okuyungibwa mu butonde nga tewali hemolysis.

2 Funa serum egenda okugezesebwa emirundi 10 mu serum dilution plate (okugeza:Add 15UL ya serum okugezesebwa ku 135UL sample diluent).

Weetegereze: Ebifuga ebitali birungi n’ebirungi tebyetaagisa kufukibwa.

1.3 Enkola y’okugezesa .

1 Twala microplate ya enzyme (okusinziira ku muwendo gwa samples, esobola okukutulwamu n’ekozesebwa emirundi egiwerako), ssaako 100UL ya diluted serum ku enzyme label microplate, oteeke enzizi bbiri buli emu for negative control ne positive control, era osse 100ul ku reaction luzzi. Sampuli ekankanya mpola mu luzzi, ng’okozesa firimu y’okusiba okusiba okubikka ku ssowaani, n’okufumbira ku diguli 37 okumala eddakiika 30.

5 Ggulawo firimu y’ekyuma ekisiba, ssika ekizimbulukusa mu luzzi lw’ekyuma, ssaako 300μl ez’amazzi ag’okunaaba mu kusengejja okukola mu buli luzzi, n’oluvannyuma amazzi ogasuule mu luzzi. Ddamu emitendera egyo waggulu emirundi 5, era okusembayo osale okale ku lupapula oluyonjo olunyiga.

3 Okwongerako 100μl enzyme conjugate ku buli luzzi, bikka ku pulati, era ofumbire ku 37℃okumala eddakiika 30.

4Ggulawo firimu y’ekyuma ekisiba, sengeka ku luzzi lw’ekisengejja, ssaako 300μl ez’amazzi ag’okunaaba mu kusengejja okukola ku buli luzzi, n’oluvannyuma amazzi ogasuule mu luzzi. Ddamu emitendera egyo waggulu emirundi 5, era okusembayo osale okale ku lupapula oluyonjo olunyiga.

5 Okwongerako 100μl substrate solution ku buli luzzi, bikka ku pulati n’ofumbira okumala eddakiika 10 ku 37℃okuva ku kitangaala.

5 Okwongerako 50μl stop solution ku buli luzzi, era osalewo ekivaamu mu ddakiika 5.

 

Eddaala 2 Okuvvuunula ebivuddemu .

Omuwendo gwa OD450 ogwa buli luzzi gwasomebwa ku 96-well microplate reader.

2.1 Okukakasa .

Omuwendo gwa OD450NM ogwa wakati ogwa negative control gwali < 0.2 era omuwendo gwa OD450NM ogwa wakati ogw’okufuga okulungi gwali munene okusinga oba gwenkana 0.7, ebyava mu kugezesa byali bituufu. Bwe kitaba ekyo, okugezesebwa kulina okuddibwamu.

2.2 Enkola y’okubalirira .

Ku buli sampuli, bala S/ P nga bwe kiri wansi ng’okozesa sampuli erongooseddwa n’emiwendo gy’okufuga:

2.3 Okuvvuunula ebivuddemu .

Bwe S/P < 0.2, esalirwa omusango nga negativu.

S/P > 0.3 bw’eba esaliddwawo nga nnungi.

Bwe 0.2 ekitono oba ekyenkana S/P ekitono oba ekyenkana 0.3, kisalibwawo ng’ekibuusabuusa, era ne kiwa amagezi okuddamu okutwala sampuli n’okugezesa oluvannyuma lwa wiiki emu.

 

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