Sar-Cov-2 Ho Nyanela Li-Antibody tse Antibody Ho Antiction (Elisa)
【Molao-motheo oa】
Kit-Cov e nyenyefatsa li-antictics tse anngoeng ho ipapisitse le mokhoa oa ho matlafatsa elisa.
U sebelisa Receptor Contptor Recing Domain (RBD), protheine e tsoang protheine ea viral spike (s) ea seleng
Receptor ACE2, tlhahlobo ena e etselitsoe ho etsisa tšebeliso ea liphoofolo tsa vaerase ea ho sitisa koluoa ea baeti.
Li-calibration, taolo ea boleng le serum kapa lipelaelo tsa plasma li tsoakane ka botsomi
Buffer e nang le Hace2-HRP e kopanya e rarolloang ka li-tubens tse nyane. Ebe metsoako e fetisetsoa ho
Li-gorkante li na le li-swebinant li-skminant smobinilinant stom-mb-2 rbd sekhechana (rbd) bakeng sa
incubation. Nakong ea poloko ea metsotso e 30, RBD e ne e le antibolt e khethehileng ea libetsa, QC le
disampole li tla qothisana lehlokoa le Hace2-Hrp bakeng sa ho hokahanya rbd e sa khoneheng lilibeng. Kamora
Ho inchutation, liliba li hlatsuoa makhetlo a 4 ho li tlosa ho se tsebe letho2-HRJugate. Tharollo ea
TMB ea eketsoa mme ea kenngoa metsotso e 20 ka mocheso oa kamore, e bakang nts'etsopele ea a
'mala o moputsoa. Nts'etsopele ea mebala e emisoa ka tlatsetso ea 1n HCL, 'me ho abeloa ke
e lekantsoe le Spectrophotometrically ho 450 nm. Botebo ba 'mala bo thehiloe ho lekana le
Palo ea enzzyme e teng, 'me e amana hantle le palo ea litekanyetso tse anngoeng ka tsela e tšoanang.
Ka ho bapisoa le curve ea likhatiso e thehiloeng ke likatse tse fanoeng, tse shebaneng
Ho sitisa li-antibodies tse sa lumellaneng tsa sampole e sa tsejoeng ka nako eo e baloa.
【Lisebelisoa tse hlokahalang empa ha li fanoe】
1. Metsi a khethiloeng kapa a leketlile
2. Likopi tsa pripete: 10μl, 100μl, 200μl le 1 ML
3. Malebela a pepeli
4. Mang ea Microplate Berter o khona ho bala ho bala 450nm.
5. Pampiri e amohelang
6. Param ea graph
7. Vortex Mixer kapa e lekanang
【Pokello ea phatlalatso le polokelo】
1. Serum le li-Sampole tsa plasma li bokelloa ka li-tubes tse nang le K2-Edta li ka sebelisoa bakeng sa k2-Edta e ka sebelisoa bakeng sa kit.
2. Liphetoho li lokela ho akheloa 'me li ka bolokoa nako e ka bang lihora tse 48 ka 2 ° C - 8 ° C ho abela.
Lipapatso li ne li tšoaretsoe nako e telele (ho fihlela likhoeli tse 6) e lokela ho ba leqheka hang ho assa le.
Qoba ho pheta-pheta hangata-nyaka.
Protocol
【Boitokisetso bo ntseng bo ntlafala】
1. Libaka tsohle li lokela ho nkuoa ho tloha sehatsetsing mme li lumelletsoe ho khutlela mochesong oa kamore pele li sebelisoa
(20 ° ho 25 ° C). Boloka linkhoe tsohle ka sehatsetsing hang hang kamora ho sebelisoa.
2. Li-dempole tsohle le lits'ila li lokela ho ba tse halikiloeng pele li sebelisoa.
3. Hace2-hrp tharollo ea tharollo: Dulitution Hace2-HRP
Buffer. Mohlala, Dulution 100 μl of Hace2-HRP
Etsa tharollo ea Hace2-HRP.
4. 1 "
Karolelano ea mofuta oa molumo oa 1:19. Mohlala, Delute 20 ml ea 20 × a hlatsoang tharollo e nang le 380 ml ea deione e entsoeng kapa
Metsi a hlokofalitsoeng a etsa tharollo ea 400 ml ea 1 × af.
【Ts'ebetso ea Teko】
1. Ka lehlakoreng le leng, aliquot 120μtse tharollo ea Hace2-HRP.
2. Kenya 6 μl ea li-caligram tse 6
3. Fetisetsa 100μl ea motsoako o mong le o mong o lokiselitsoeng mohatong oa 2 ho ea li-greents tse lumellanang tsa microplate ho latela
ho tsepamisoa pele ho fana ka tumello.
3. Koahela poleiti e nang le senotlolo sa poleiti le incubte ho 37 ° C bakeng sa metsotso e 30.
4. Tlosa senotlolo sa poleiti 'me u hlatsoe poleiti e ka bang 300 μl ea 1 × b.l
5. Tlanya poleiti ka thaole ea pampiri ho tlosa mokelikeli o setseng ho lilibeng kamora mehato.
6. Kenya 100 μl ea Tharollo ea TMB ho e 'ngoe le e' ngoe e 'ngoe e tsoekere lefifing le lefifing ho 20 - 25 ° C bakeng sa metsotso e 20.
7. Kenya 50 μl ea tharollo ea ho emisa sebakeng se seng le se seng ho emisa karabelo.
8. Bala ho ba teng ka microplate 'mali ho 450 nm nakong ea metsotso e 10 (630Nm e le ea kenyelletso ea
khothalletsoa ts'ebetso e phahameng ea ho nepahala).