??? 1.????????(ELISA) 2.??????

1
??? 1.????????(ELISA)2.??????
2
1.????????(ELISA)
3
??????

• ??
• ???-????????????,???????????????????????,?????
  ??,?????????????
• ??
• ???????131I,32P
• ??????FITC,PE
• ??????HRP,AP

4
??????(Enzyme ImmunoAssay,EIA)

• ????????????????????
• ???????(HRP),?????(AP)
• ??
• ?????????????????
• ???????????????,pg??????
• ??????????????????
• ??
• ????????????????
• ?????? ????????????

5
????????(Enzyme-Linked ImmunoSorbent Assay,ELISA)
1.??
?????????????(??????)????????????
2.??

• ???(Indirect ELISA)
• ????????????,?????
• ?????????
• ??????(Sandwich ELISA)
• ????????????,?????
• ???????????
• ???(Competitive ELISA)
• ???????????????,????????
• ??????????????

6
????????(Enzyme-Linked ImmunoSorbent Assay,ELISA)
3. ??(????ELISA??)
??
7
???? ???????? ????
8
????

• ???????? ??
• ?????????(???) ?????
• HRP?????IgG ??
• ?????0.05M pH9.6??????
• ????0.05 Tween 20?0.01M pH7.4 PBS
• ????? pH5.0?????????
• ????OPD 10mg,?????25ml,30 H2O2 40µL
• ???,???

9
????
??????
?3mL N.S,??
2000rpm,5min
1.?????
?3mL N.S,??
2000rpm,5min
?2mL?????
??2mL???,???????
?????????1400??
????,??100µL/???????
2.????
4?,???,18h
10
????
???????????(4?/?),??????
????(200µL/?)??3?,3min/?
1
2
3
4
3.?????
????,??????100µL/?
??
??
??
??
37?,???,30min
??????
4.???
????(200µL/?)??3?,3min/?
????????100µL/?
37?,???,30min
??????
5.??
????(200µL/?)??3?,3min/?
??????100µL/?
6.????
????,10min
11
????

• ???????
• ????

1 2 3 4
??
??
??
??
12
????

• ????,???????
• ?????,??????
• ????????????

13
2.??????(Complement fixation Test, CFT)
14
??

• ??????
• ?????????,??????(SRBC)????(?SRBC???)???????
  ???????

15
??

• ?????????????
• ??????Ag?Ab???Ab?Ag?????Ag-Ab??????????
• ??????????????

16
1No Ab 2Ab
???? ??????
???? SRBC?SRBC??
??
1
2
1
2
Step 1 ??????????
Step 2 ????????
Step 3 SRBC???
17
Ab ??
No Ab ??
Ag
Ag
Ag
Ag
Y

18
??

• ?????????
• ????56?,30min
• ??????
• 2?????
• ???

19
??
?? ???
?? ???
?? ???
SRBC ???
???
1
2
3
4
5
(1?/2?)



0.2mL
????
0.2mL
??



0.2mL
0.2mL
??

0.2mL
0.2mL
0.2mL
0.2mL
N.S

0.2mL
0.4mL
0.8mL
0.2mL
??,37?,??15min

???
0.2mL
0.2mL
0.2mL
0.2mL
0.2mL
0.2mL
SRBC
0.2mL
0.2mL
0.2mL
??,37?,??15min
20
????
1.??????????????,?????????????
??
???
2.
??? ????? ????? ????? SRBC???
? ?? ?? ?? ???
21
????

• ????????????,???????
• ????????????