Ukucaciswa
Izinqubo Zokuhlola Nokuhumusha
ISINYATHELO 1 Inqubo Yokuhlola
1.1 Ukulungiselela Isixazululo Sokugeza
Uketshezi lokugeza lokugxilisa okusebenzayo : Ngaphambi kokusetshenziswa, isisombululo sokugeza kufanele sibuyiselwe ekamelweni lokushisa (20-25 degree) futhi sinyakaziswe ngokulinganayo ukuze senze okufanayo. Bese uhlanza izikhathi ezingu-20 isisombululo sokugeza esigxiliwe ngamanzi ahlanzekile (isibonelo, isixazululo sokugeza esigxilile esingu-5ml sengezwa ku-95ml amanzi ahlanzekile), hlanganisa kahle, futhi uketshezi lokugeza oluhlanjululwe lungagcinwa ku-2-8℃izinsuku ezingu-7.
1.2 Ukulungiselela Isampula
① Igazi lesilwane lonke lathathwa, lafakwa i-centrifused ku-4000rpm imizuzu engu-10, futhi kwaqoqwa amandla amakhulu. I-Serum ingabuye ihlanganiswe ngokwemvelo ngaphandle kwe-hemolysis.
② Nciphisa isampula ye-serum 1:25 (engeza isampula ye-serum engu-5 ku-120ul yesampula yokuxubeka) bese uxuba kahle.
Qaphela: Izilawuli ezingezinhle nezihle akumele zihlanjululwe.
1.3 Ukulungiselela i-enzyme conjugate
Nciphisa i-enzyme conjugate 1:25 (engeza i-5ul enzyme conjugate ku-120ul enzyme diluent) bese uxuba kahle.
1.4 Inqubo Yokuhlola
①Thatha ilebula ye-enzyme ye-Microplate (ngokwenani lamasampuli, ingahlakazwa futhi isetshenziswe izikhathi eziningana), engeza i-100ul ye-serum ehlanjululwe kulebula ye-enzyme ye-Microplate, usethe imithombo emibili ngayinye yamasampuli angalungile namasampula, bese wengeza i-100ul ngayinye ngezilawuli ezingezinhle nezihle ekuphenduleni kahle ngokulandelanayo. Gcoba ngobumnene isampula emthonjeni, usebenzisa ifilimu yepuleti lokuvala ukuvala ipuleti, bese ufukamela ku-37℃imizuzu engu-30.
②Vula ifilimu yepuleti lokuvala, vuthulula isixazululo epuletini kahle, engeza u-300µl woketshezi lokuwasha lokugxilisa okusebenzayo emthonjeni ngamunye, bese ulahla uketshezi emthonjeni. Phinda lezi zinyathelo ezingenhla izikhathi ezi-5, bese ekugcineni ubambe ephepheni elihlanzekile elimuncayo.
③Engeza u-100µl we-Enzyme Conjugate emthonjeni ngamunye, vala ipuleti, futhi ufukamele ku-37℃imizuzu engu-45.
④Vula ifilimu yepuleti lokuvala, vuthulula isixazululo epuletini kahle, engeza u-300µl woketshezi lokugeza okugxilile osebenzayo emthonjeni ngamunye, bese ulahla uketshezi emthonjeni. Phinda lezi zinyathelo ezingenhla izikhathi ezi-5, bese ekugcineni ubambe ephepheni elihlanzekile elimuncayo.
⑤Engeza isixazululo se-substrate esingu-100µl emthonjeni ngamunye, vala ipuleti bese ufukamela imizuzu engu-10 nge-37℃kude nokukhanya.
⑥Engeza isisombululo sokumisa esingu-50µl emthonjeni ngamunye, bese unquma umphumela phakathi nemizuzu emi-5.
ISINYATHELO 2 Ukuhunyushwa komphumela
Inani le-OD450 lomthombo ngamunye lafundwa kumfundi we-microplate we-96-well.
2.1 Ukuqinisekisa
Isilinganiso senani le-OD450nm lokulawula okungekuhle kwaba<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 Indlela yokubala
Kusampula ngayinye, bala i-S/P ngendlela elandelayo usebenzisa isampula elilungisiwe kanye namanani okulawula:
S / P=(ODISAMPELA -ODNC ) / (ODI-PC -ODNC )
2.3 Ukuhunyushwa Kwemiphumela
Lapho i-S/P<0.15, it is judged as negative.
Uma i-S/P Inkulu noma ilingana no-0.25, kuthathwa ngokuthi iphozithivu.
Lapho 0.15 Ngaphansi noma kulingana nokuthi S/P<0.25, it is judged as suspicious, and suggest to repeat sampling and test one week later.
Amashadi ashisayo: uhlobo lwe-porcine circovirus-ⅱ ikhithi yokuthola i-antibody elisa, uhlobo lwe-China porcine circovirus-ⅱ abahlinzeki bekhithi yokutholwa kwe-antibody elisa, abakhiqizi, imboni
