to3methods
I
directeLIsAK
ow
Age
zymelabeledseco
daryAbtodetectu
k
ow
Absa
dwicheLIsAa
dcompetitiveeLIsA
Firstthek
ow
a
tibodyora
tige
isfixedo
asolidcarrierThe
asamplea
dthecorrespo
di
ge
zymeli
kedAbAgareaddedtobi
dtotheAgAbattachedtothesolidcarrierTheresulti
gAgAbcomplexa
dtheexcessAbAgareseparatedbywashi
gFi
allythesubstrateisaddeda
dcatalysiscauseacolorcha
gewhichca
bemeasuredeLIsAhastheadva
tagesofhighse
sitivitya
dspecificityItcombi
esthespecificityoftheAbAgreactio
withthecatalysisofe
zymes
methods
f1preparatio
ofAgadd3ml
stotubeco
tai
i
gdefibratedsRbca
dmixThe
ce
trifuge2000rmp5mi
add3ml
stothetubea
dmixedthe
take2mlpackedsRbcAdd2mlDDwa
dshattersRbca
ddilutewithcoati
gbufferi
aratioof1400
2coati
gAgAdd100μLofAgtoeachwellofeLIsAplatethe
use
stowashforfivetimes
3preparetheserumremovetheeyeballofthesRbcimmu
izedmicea
dcollectthebloodi
toeptubewaitforabout5mi
utethe
ce
trifugeat12000rmpfor10mi
takethesuper
a
ta
ddilutetheserum110
4Addtestserumsig
a
dadd100μLofsolutio
toeachwellthe
putiti
ahumidifiedboxu
der37℃for45mi
5Addseco
daryAbDiscardthesolutio
i
eLIsAplatea
dwashtheplate3timesThe
add100μ
LofhRplabeledseco
daryAbtoeachwella
dputiti
ahumidifiedboxu
der37℃for30mi
f6showi
gcolorDiscardthesolutio
i
eLIsAplatea
dwashtheplate3timesThe
add100μLofsubstratesolutio
toeachwellshowcolori
darkfor10mi
result
the
o1wellisbla
k
o2is
egativewell
o3ispositivewell
o4istestwellThe
o3a
d
o4becameyellowwhile
o1a
d
o2iscolorlessThecolorofthe
o4wellisdeepertha
the
o3
a
alysis
Theresulti
o4wellispositivebecauseitbecomeyellowwhichmea
stherearea
tibodiesi
the
o4wellalsoi
dicatesthatsRbc
immu
izedmicehadproduceda
tibodiesThecoloroftestwell
o4wellisdeepertha
thepositiveco
trolwell
o3wellsoa
tibodiesamou
ti
testserumismoretha
thati
thepositiveco
trol
Test3hyperse
sitivityTestofgui
eapig
pri
ciples
fhyperse
sitivityreferstotheselftissuei
jurya
dorbiologicaldisorderwhe
these
sitizedi
dividualco
tactsthesameallerge
agai
hyperse
sitivityistheab
ormalorpathologicimmu
erespo
seAccordi
gtothemecha
ismsofhyperse
sitivityitcouldbedividedi
tofourtypestypeⅠ、Ⅱ、Ⅲa
d
ⅣTypeIhyperse
sitivityisalsocalledimmediatetypehyperse
sitivityora
aphylaxisThea
tige
leadstohyperse
sitivityiscalledallerge
Thea
tibodytakeparti
a
aphylaxisismai
lyIge
Allerge
i
ducedperiod
I
typeIhyperse
sitivityallerge
e
tersbodyforthefirsttimea
dstimulatesplasmacellstoproduceIgeIgecouldbi
dtothecellsurfaceFcεRofthemastcellsorbasophils
Allerge
stimulatedperiodwhe
thesameallerge
e
tersthese
sitizedbodyagai
allerge
couldbi
dtotheIgeo
thecellsurfaceThe
aseriesofreactio
happe
whichleadtothecellactivatio
a
dbiologicalmediatorsbei
greleasedfromthesecellsThesemediatorsleadtotheattackoftypeIhyperse
sitivity
effectperiod
fhistami
eisthemai
mediatorthathasimporta
tbiologicaleffectsTherapidreleaseofhistami
ecouldi
creasevascularpermeabilir