1.What is a microarray

1
??????????

• ????????????
• ?????? ???

2
1.What is a microarray

• Microarray ? Biochip (????)???????????????????????
  ??????????????,???????????,???????????????????????
  ?????????????????
• A microarray is an ordered array of microscopic
  elements on a planar substrate that allows the
  specific binding of genes or gene products.

3
1.What is a microarray

• Four distinct characteristics of microarray
• Microscopic target elements or spots
• Planar substrates
• Rows and columns of elements
• Specific binding between microarray target
  element on the substrate and probe molecules in
  solution.

4
1.What is a microarray

• Which is the probe?and
• which is the target?
• There are some differences between China and
  United States of America.
• In U.S.A. the probe are usually in solution.
• In China the probe is a reagent to detect target
  from sample.

5
Biochips?????

• ????? Protein chips(protein microarray)
• ???,????
• ????? Gene chips(Gene microarray)
• ?DNA??,?????
• ??????Laboratory-on-a-chip
• or Microfluidics-based chip

6
2.Microarray manufacturing

• The three main manufacturing methods
• Contact printinguses direct contact between the
  printing implement or the sample contained in the
  implement and the microarray substrate
• Noncontact printingon-the-fly printing including
  ink-jet tech.,microsolenoid tech.,and thermal
  bubble jet dispenser
• Semiconductor tech photolithographic masks and
  micromirrors.

7

• Microscopic elements
• ?Semiconductor tech produce 1530µm features
• ? Printed spot size is 50350 µm
• ? Tissue spot is 200600 µm
• Macroarrays contain sample spot sizes of about
  300 microns or larger
• microarray are typically less than 200 microns in
  diameter

8
Gene chips???

• ? ????????
• O d NTP- 5-??????

??
O dNTP-OH dNTP -5-????
9

?????????
10
Gene chips???

• cDNA???
• ?????? Silanized slide(?DNA?????)
• Silane slide
• ?????? Silylated slide(UV???DNA??)
• Polylysine slide
• ???????

11
Nytran???

• ?????
• ??????10.4N NaOH/10mM EDTA,
• ???????????Nytran????,
• ?120mj/cm2UV????(Stratalinker 1800,
• Stratagene)
• ?????
• PCR???????(???)?Nytran???
• ???,?0.4N NaOH/3M NaCl???5???
• ?DNA,?????6SSC??5????,?
• ?120mj/cm2UV?????

12
???????????????????????
13
?????
14
????????????Cy3-dNTP?Cy5-dNTP???????????DNA??????
,??????????,???????????????????
?????
15
????????
16
????????
17
Gene chips???

• ? ??????
• ? ?????????
• ? ????
• ? ?????
• ? ??
• ? ??????
• ? ???????
• ? HLA??

18
(No Transcript)
19
 
????????????????
ATACGTTA  
TACGTTAG  
 
ATACGTTA  
TACGTTAG  
?????  
ACGTTAGA  
CGTTAGAT  
GTTAGATC  
ACGTTAGA  
CGTTAGAT  
ATACGTTAGATC  
???????  
GTTAGATC  
TATGCAATCTAG  
???  
??????????

20
???????

• 1.????????(UV)??????

21
(No Transcript)
22

• 2. ??????????
• ??HRPO?AKP
• ????
• ????Cy3?Cy5?FITC?RB200??????

23

3.???????????
??????
POTOMAC
24
??????
POTOMAC
25
??????-???
POTOMAC
26
??????
POTOMAC
27
??????????
POTOMAC
28
????
??????????
29
?????????3???A,?????????,?????????
30
??????,????1??,?????100µl?
31
????????,???6???B?
32
???B?????,??10???C?
33
???C?????,????6???D?
34
?????0.5???,??????????????????
35
???PBT-X2-02????????????
36
Protein chip???

• ??????-???????????????????
• ????????
• ? HLA??
• ? ?????
• ????????

37
?????????? ?????????? ?????LAM
?16KDa?38KDa?? LAM???????,??????????????? 38KDa
?????????10??? 16KDa??????????????(?????????)?,
??????(MOTT)???? ??????????93
38
?????????????(????LAM)

• ??????(Mycobacterium tuberculosis)????????(LAM)???
  ????????????LAM???????????????,??????????????????L
  AM????????????????
• LAM?????????,???????????
• ???????
• 1)   ???
• 2)   ??T??????
• 3)   ????????????
• 4)   ????????
• 5)   ??????????
• ??,LAM?????????????????????,??????????????
  ?,?????????

39
??????MPT63??(16KDa)

• ???MPT63??????????,????16kDa,????MPT63,??159???(aa
  )????,29aa????,130aa??????MPT63?????????????????MP
  T63????????????(M tuberculosis complex,
  MTC)?MPT63?????????(M avium)????????????
• MPT63????????MTC??????MPT63,??MPT63???MTC????????
  ???MPT63??????MPT63,???????,?????PPD??,????????PP
  D???????,?????ELISA,MPT63??MPT63?????????????PPD??
  ??,????????PPD?BSA(????)??,??,MPT63???????????????
  ?,??????????????????

40
???????38kDa??

• ???????,?????????5(Antigen 5),?????????78(Antigen
  78)?Coates?????????????????38kDa,????38kDa??
• 38kDa ??????,?????? ,?25?????,???????,???????3820
  0Da(????),?????????????????? ?
• 38kDa????????????(crossed immunoeleltrephoresis,C
  IE)????????????38kDa??,?????????????
• ????????????????(M boris)??38kDa??,???????????????
  ??10? ?
• ?-BCG???????38kDa???? ,?BCG??38-kDa?????? ?
• ??????38kd?????100,???????MOTT???????????????

41
TB????????
2
1
4
2
4
1
2
4
1
3
3
3
LAM 16KDa 38KDa H2O
1
???? ???
2
3
4
42
?????????DIGFA??(????)???????

• ?????????????1285???,???326?,???769?,?????1095????
  ???85.21? 
• ????????????????????????????????
  ??????????? (????)

43
?????????LAM?16?38kD???????

• ???????????????????
• ??????????????
• ????????
• ???????????
• ????????????????

44
??????AMI????

• ????(Myoglobin)???????????
• 1-2????? 4-8?????? 20-36??????
• ???????(CK-MB)???AMI???????
• 3-8????? 8-24?????? 3?????
• ????I(Troponin I)?????????????
• 4-8????? 12-16?????? 5-10?????
• ??C????(hs-CRP)
• 4-8????? 12-16?????? 5-10?????

45
??????(AMI)??????????????????????????
??????????
? ? ???? ??? ?? ????? ??rc??
CK-MB ??ECL2010 524 0.852
MB Bayer??ACS 180plus 524 0.915
cTnI Bayer??ACS 180plus 524 0.915
hs-CRP ??? Immage 524 0.89
46
??STD??????

• ?????UU???(?1,3,6,14??????)????(?2,4,5,7,8,9,10,11
  ,12,13??????)????,??????????????
• ?????CT???????(DK?)???????? ( LGV)????
• ???UU/CT???85???
• ??????UU/CT???1/3?

47
??STD????????1.???????(????), 2.?????????,
3.????????, 4.???????????????????????
?? ? ? ?? ???() ???()
CT ???? 955 88.1 80.7
UU ????? 955 86.1 81.2
48
??TORCH????

• ??TORCH??(IgM?IgG)
• ???????????????????????(???)???
• ??????,????????
• IgM??????????,IgG???????????

49
TORCH???????

• ??30-46?????????????,?????????,????,????????????
  ,??,???????????
• ?????????,???????????????????????????????????????,
  ???????????

50
TORCH???????

• ??CMV??????????????,??????????????????????????????
  ???,?????????????????????????,????????????,???????
  ??
• ????HSV???????????,??????????,?????????,?????,????
  ,???????????

51
??????(HP)IgG??????

• ??Hp??????????????A(CagA) ????(Ure)??????(HSP)
• ????
• ??HP????????????A(CagA)?????????????????,?CagA??,
  ??CagA?????????????????????????
• ????????HP??????(Ure),Urc??????Hp??????
• HSP???????, HP?HSP????????,??HP????????,?????????
  ,????HSP60?????????????

52
??????????

• ????????
• ??dsDNA,Histone,SSA/Ro,SSB/La,Sm,RNP,Jo-1,Scl-70,
  Rib-P?????
• ?????????????????????????????????????????????????
  ??????????????

53

• ???????????? 20031128
• ?????
• C12?????? ????
• ?????????? ?????
• HP???? ??
• UU CT HPV???? ??
• ?????????

POTOMAC
54

• ???????????? 20031128
• ?????
• ???????????????
• ??????????
• AMI????????(???????) ??
• UU CT???? (???????) ??
• TB?????? (???????,??????) ??
• SARS????
• TORCH???? (???????) ??
• ???????????? (???????) ??

POTOMAC
55

• ???????????? 20031128
• ?????
• HLA-DR??????
• HLA-AB??????
• SARS??????????
• ?????????
• SARS????
• TB?????? ??
• ???????? ??

POTOMAC
56

• ?????????
• a. ???????????????
• b. ???????????????????
• ??????,???,???
• ????????????????????????????????????
  ??????????????????

57
POTOMAC
58
(No Transcript)
59
?????

• ????????
• ????
• ????????(?????????????)
• ???????????????????(????????????????????????)

60

• THANK YOU FOR
• YOUR ATTENTION !