Kokoro Ẹsẹ Ati Ẹnu -O Antibody Elisa Kit

Sipesifikesonu

FMDV-Ìwọ Ab Elisa

Lilo ti a pinnu: Kokoro Ẹsẹ Ẹsẹ ati Ẹnu -O

Apo Iwari ELISA Antibody jẹ fun ipinnu Ẹsẹ ati apakokoro ẹnu ni omi ara bovine ni vitro. Idanwo naa ni a lo lati ṣe ayẹwo ẹsẹ ati ẹnu arun ọlọjẹ iru O antibody ajesara ti awọn oko.

Apeere: omi ara

Awọn ohun elo ti a beere Ṣugbọn Ko Pese:

Omi ti a wẹ

Pipettor

Oluka Microplate 96 daradara (450nm)

Aago

Centrifuge (4000rpm)

Ẹrọ fifọ

Lọla (iwọn 37)

Awọn ilana Igbeyewo & Itumọ

Igbesẹ 1 Ilana Igbeyewo

1.1 Wẹ Solusan Igbaradi

Wifọkansi orking womi eeru:Ṣaaju lilo, ojutu fifọ yẹ ki o tun pada si iwọn otutu yara (iwọn 20-25) ati gbigbọn paapaa lati jẹ ki o jẹ aṣọ. Lẹhinna fi omi ṣan ni igba 20 ojutu ifọkansi pẹlu omi ti a sọ di mimọ (fun apẹẹrẹ, ojutu ifọkansi 5ml ti wa ni afikun si omi mimọ 95ml), dapọ daradara, ati omi fifọ ti fomi le wa ni ipamọ ni iwọn 2-8 fun awọn ọjọ 7.

1.2 Apeere Igbaradi

① Gbogbo ẹjẹ ẹranko ni a mu, ti a fi si 4000 rpm fun awọn iṣẹju mẹwa 10, ati pe a ti gba supernatant. Omi ara le tun ti wa ni coagulated nipa ti lai hemolysis.

② Dilute omi ara 1: 25 (fi 5ul serum sample to 120ul sample diluent) ati ki o dapọ daradara.

Akiyesi: Awọn iṣakoso odi ati rere tun nilo dilute.

1.3 Enzyme conjugate Igbaradi

Dilute awọn enzymu conjugate 1:90 (fi 5ul enzymu conjugate (x100) to 445ul sample diluent) ati ki o dapọ daradara.

1.4 Igbeyewo Ilana

① Mu aami enzymu Microplate (ni ibamu si nọmba awọn ayẹwo, o le ṣajọpọ ati lo ni ọpọlọpọ igba), ṣafikun 100ul ti omi ara ti a ti fomi si aami Microplate, ṣeto awọn kanga meji kọọkan fun odi ati awọn apẹẹrẹ rere, ati ṣafikun 100ul kọọkan fun odi ati awọn iṣakoso rere si iṣesi daradara lẹsẹsẹ. Fi rọra gbọn awọn ayẹwo ni kanga, lilo awọn lilẹ awo fiimu lati bo awo, ati incubate ni 37 iwọn fun 30 iṣẹju.

② Ṣii fiimu awo edidi, gbọn ojutu ti o wa ninu awo daradara, ṣafikun 300µl ti omi ifọkansi iṣiṣẹ ṣiṣẹ si kanga kọọkan, lẹhinna sọ omi bibajẹ ninu kanga naa. Tun awọn igbesẹ ti o wa loke ṣe ni igba 5, ati nikẹhin gbẹ gbẹ lori iwe ifamọ ti o mọ.

Fi 100µl ti Enzyme Conjugate sinu kanga kọọkan, bo awo naa, ki o si fi sii ni iwọn 37 fun awọn iṣẹju 45.

④ Ṣii fiimu awo lilẹ, gbọn ojutu ninu awo daradara, ṣafikun 300µl ti omi ifọkansi iṣiṣẹ ṣiṣẹ si kanga kọọkan, lẹhinna sọ omi bibajẹ ninu kanga naa. Tun awọn igbesẹ ti o wa loke ṣe ni igba 5, ati nikẹhin gbẹ gbẹ lori iwe ifamọ ti o mọ.

Ṣafikun ojutu sobusitireti 100µl si kanga kọọkan, bo awo naa ki o si fi sii fun iṣẹju mẹwa 10 ni iwọn 37 kuro ni ina.

⑥ Ṣafikun ojutu iduro 50µl si kanga kọọkan, ki o pinnu abajade laarin awọn iṣẹju 5.

Igbesẹ 2 Abajade Itumọ

Iye OD450 ti kanga kọọkan ni a ka lori oluka microplate 96-daradara.

2.1 Ifọwọsi

Iwọn OD450nm ti iṣakoso odi jẹ<0.15 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 Iṣiro Ọna

Fun apẹẹrẹ kọọkan, ṣe iṣiro S/P gẹgẹbi atẹle nipa lilo ayẹwo atunṣe ati awọn iye iṣakoso:

2.3 Abajade Itumọ

Nigbati S/P<0.15, it is judged as negative.

Nigbati S/P Ti o tobi ju tabi dogba si 0.25, o jẹ idajọ bi rere.

Nigbati 0.15 Kere ju tabi dogba si S/P<0.25, it is judged as suspicious, and suggest to repeat sampling and test one week later.

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