Mtundu wa Circovirus wa Nkhumba-II Chipangizo chakumanyira ELISA
Nambala ya chiyelezgero: VD510E
Ab Elisa
Chiyelezgero: seramu
Kulongosora
Kulongosora
PCV - II Ab Elisa
Kugwiriskira Ntchito:
Mtundu wa Circovirus ya Nkhumba -II Chipangizo cha ELISA chikumanyira chitetezo cha PCV mu seramu ya nkhumba mu vitro. Mayeso agha ghakugwiriskira nchito kuti ŵawone awo ŵapokera katemera kufuma ku katemera wa mtundu wa porcine circovirus-II wa mafamu gha nkhumba.
Chiyelezgero:seramu
Vinthu ivyo vyapelekeka:
Ndondomeko ya mayeso na kumasulira
STEP 1 Nthowa yakuyezgera
1.1 Kunozga Nthowa Yakuchapira
Kugwira ntchito kwa maji ghakuchapira: Pambere mundaseŵezeske, maji ghakuchapira ghakwenera kuwelera ku msinkhu wa chipinda (20-25℃) na kusunkhunyika mwakuyana kuti ghaŵe ghakuyana waka. Pamanyuma pake, ŵikani maji ghakutozga ghakukwana 20 (mwachiyelezgero, 5ml ya maji ghakutozga ghakusazgapo 95ml), sakanizgani makora, ndipo maji ghakutozga ghangasungika pa 2-8℃kwa mazuŵa 7.
1.2 Kunozga kwa chiyelezgero
Ndopa zose za nyama zikatoleka, zikaŵikika mu centrifuge pa 4000rpm kwa maminiti 10, ndipo ndopa izo zikaŵa pa mtunda zikatolekanga. Seramu nayo yingaŵa yakukhora mwachibadwa kwambura hemolysis.
Kuchepeska seramu 1:25 (kuwonjezera seramu 5ul ku 120ul sampulo diluent) na kusakanizga makora.
Ziŵani kuti: Vinthu ivyo vikulongozga viheni na viwemi vikwenera yayi kuchepeskeka.
1.3 Kunozga vyakupangika na enzyme
Kuchepeska enzyme 1:25 (kuwonjezera enzyme 5ul ku 120ul enzyme diluent) na kusakanizga makora.
1.4 Nthowa yakuyezgera
1Torani chimanyikwiro cha enzyme Microplate (kuyana na unandi wa masampulo, chingagaŵika na kugwiliskirika ntchito kanandi waka), ŵikanipo 100ul ya seramu yakuchepeskeka ku chiziŵizgo cha enzyme Microplate, ŵikanipo visimi viŵiri chilichose cha masampulo ghaheni na ghawemi, ndipo ŵikanipo 100ul kwa masampulo ghaheni na ghawemi. Kugwedezga sampulo mu chisimi mwakuzika, kugwiriskira ncito filimu ya mbale yakusindikizga kuti yibenekelere mbale, na kuŵika pa 37℃kwa maminiti 30.
2Julani filimu ya mbale yakusindikizga, kugwedezga yankho ilo lili mu chisimi cha mbale, ŵikanipo 300μl ya maji ghakusambira ghakugwira ntchito ku chisimi chilichose, ndipo pamanyuma mutaye maji agho ghali mu chisimi. Bwerezganipo ndondomeko za pacanya apa maulendo 5, ndipo paumaliro muwume pa pepala lakutowa lakupokera maji.
3Sakanizgani 100μl ya Enzyme Conjugate ku chisimi chilichose, vikani mbale, na kuŵika pa 37℃kwa maminiti 45.
4Julani filimu ya mbale yakusindikizga, kugwedezga yankho ilo lili mu chisimi cha mbale, ŵikanipo 300μl ya maji ghakusambira ghakugwira ntchito ku chisimi chilichose, ndipo pamanyuma mutaye maji agho ghali mu chisimi. Bwerezganipo ndondomeko za pacanya apa maulendo 5, ndipo paumaliro muwume pa pepala lakutowa lakupokera maji.
5Sakanizgani 100μl ya substrate solution ku chisimi chilichose, vikani mbale na kuŵika mu mphindi 10 pa 37℃kutali na ungweru.
Sakanizgani 50μl stop solution ku chisimi chilichose, ndipo manyani ivyo vikufumamo mu mphindi 5.
CHIGAŴA 2 Kumasulira vyakusangikamo
Mtengo wa OD450 wa chisimi chilichose ukaŵazgika pa chipangizo chakuŵazga cha microplate cha chisimi cha 96.
2.1 Kukhozgera
Pafupifupi mtengo wa OD450nm wa kulaŵilira kwakuheni ukaŵa .<0.1 and the average OD450nm value of positive control /the average OD450nm value of negative control was ≥5, the experimental results were valid. Otherwise, the test should be repeated.
2.2 Nthowa yakuŵerengera
Pa sampulo yiliyose, ŵerengerani S/P nga umo vikuŵira mwakugwiliskira nchito sampulo yakunozgeka na vigaŵa vyakulongozga:
S / P =(ODCHIYEZGELERO-ODNC) / (ODPC-ODNC)
2.3 Kumasulira vyakufumapo
Para S/P<0.15, it is judged as negative.
Para S/P Yikuru kuluska panji yakuyana na 0.25, yikuwoneka kuti njakwenelera.
Para 0.15 Pachoko panji kuyana na S/P .<0.25, it is judged as suspicious, and suggest to repeat sampling and test one week later.
Tag yakutentha: mtundu wa circovirus wa nkhumba-I chida chakumanyira chitetezo cha elisa, China mtundu wa circovirus wa nkhumba -I chida chakuziŵizga chitetezo cha elisa ŵakupeleka, ŵakupanga, fakitare
Kumanyuma: Yayi
