Munthu uyo ​​wali na nthenda ya Parvovirus

Phafififize

 

 

Nthowa zakuyezga & Kulongosora

 

STEP 1 Nthowa Yakuyezga

1.1 Kunozga Nthowa za Wash

Wkuŵika kuŵika maghanoghano ghake pa wphulusa:Pambere mundagwiriskike nchito, nthowa yakuchapira yikwenera kuwezgeka ku kutentha kwa chipinda (20-25℃) na kusunkhunyika mwakuyana kuti yiŵe yakuyana waka. Pamanyuma pake, dilute 20 nyengo zakuchapira na maji ghakutozgeka (mwaciyelezgero, nthowa yakuchapira ya 5ml yikusazgikira ku maji ghakutozga gha 95ml), kusakanizga makora, ndipo maji ghakugezeramo ghakudumulika ghangasungika pa 2-8℃kwa mazuŵa 7.

1.2 Chiyelezgero

Nyama yiliyose yikatoreka, centrifuged pa 4000rpm kwa maminiti 10, ndipo supernatant yikasonkheka. Seramu nayo wangaŵa wakuchimbira mwachibadwa kwambura hemolysis.

② Delute kuti seramu yiyezgeke maulendo 10 mu mbale ya dilution ya serum (mwachiyelezgero:kusazgirapo 15ul ya seramu kuti yiyezgeke ku 135l sampulo diluent).

Ziŵani kuti: Vinthu ivyo vikupangiska kuti munthu waŵe na suzgo la maghanoghano vikwenera yayi kuchepeskeka.

1.3 Nthowa

Pangani chiziŵizgo cha enzyme Microplate (kuyana na chiŵelengero cha masampulo, chingagaŵika na kugwiliskirika ntchito kanandi waka), kusazgapo 100ul ya seramu yakuchepeskeka ku enzyme chikwangwani cha Microplate, kuŵika visimi viŵiri vyose kuti viŵe na mazaza ghaheni na kulaŵilira viwemi, ndipo kusazgako 100ul ku chisimi cha vyakuchitika mwakuyana. Mutembenuzge mwakuzika sampulo mu chisimi, kugwiriskira ncito filimu ya mbale yakudinda kuti yibenekelere mbale, na kuŵika pa 37℃kwa maminiti 30.

‐Jepen filimu ya chidindo cha chidindo, kugwedezga yankho mu chisimi cha mbale, ŵikanipo 300μl ya maji ghakuchapiramo ghakugwira ntchito ku chisimi chilichose, ndipo pamanyuma mutaye maji mu chisimi. Bwerezganipo masitepe agho ghali pacanya apa maulendo 5, ndipo paumaliro muwume pa pepara lakutowa lakupokera maji.

‐Add 100μl ya Enzyme Conjugate ku chisimi chilichose, kuphimba mbale, na kuŵika pa 37℃kwa maminiti 30.

‐Open filimu ya chidindo cha chidindo, sunkhunyani nthowa iyo yili mu chisimi cha mbale, ŵikanipo 300μl ya maji ghakuchapira ghakugwira ntchito ku chisimi chilichose, ndipo pamanyuma mutaye maji mu chisimi. Bwerezganipo masitepe agho ghali pacanya apa maulendo 5, ndipo paumaliro muwume pa pepara lakutowa lakupokera maji.

‐Add 100μl substrate yankho ku chisimi chilichose, kuphimba mbale na kuŵika kwa maminiti 10 pa 37℃kutali na ungweru.

‥Addd 50 μl stop trafer ku chisimi chilichose, na kusankha ivyo vikuchitika mu mphindi 5.

 

STEP 2 Kulongosoraso

 

Mtengo wa OD450 wa chisimi chilichose ukaŵazgika pa 96-well microplate wakuŵazga.

2.1 Kuwonekera

Avereji ya OD450m mtengo wa kulaŵilira kwakunanga ukaŵa <0.2 ndipo pafupifupi OD450nm mtengo wa kulaŵilira kwakukhozga ukaŵa Mukuru kuluska panji wakuyana na 0.7, vyakuchitika vyakuyezgayezga vikaŵa vyakuzomerezgeka. Kupatula apo, mayeso ghakwenera kuwerezgeka.

2.2 Nthowa yakuŵerengera

Pa sampulo yiliyose, kupenda S/ P nga ni umo vikuŵira pakugwiriskira ncito sampulo yakunozgeka na vigaŵa vyakulongozgera:

2.3 Vinthu ivyo vikupangiska kuti ŵanthu ŵaŵe na mwaŵi wa kuŵika vinthu pa malo ghanyake.

Para S/P < 0.25, yikuyeruzgika kuti ni viheni.

Para S/P > 0.3, yikupimika kuti njakukhozga.

Para 0.25 Kuchepa panji kufanana na S / P Less kuluska panji kufanana na 0.3, kukuyeruzgika kuti nkhwakukayikiska, ndipo kukupereka fundo yakuti kuwerezgapo na kuyezga sabata yimoza pamanyuma pake.

Tag yakutentha: porcine parvovivir antibody elisa kit, China parvovirus parvovirus antibody elisa kit kit, ŵakupanga, fakitare .