ĐẠI HỌC QUỐC GIA HÀ NỘI
TRƯỜNG ĐẠI HỌC KHOA HỌC TỰ NHIÊN

Trần Thị Hòa

NGHIÊN CỨU ỨNG DỤNG KỸ THUẬT POLYMERASE
CHAIN REACTION TRONG CHẨN ĐOÁN VIÊM ÂM
ĐẠO - CỔ TỬ CUNG DO Chlammydia trachomatis TẠI
BỆNH VIỆN ĐẠI HỌC Y THÁI BÌNH
Chuyên ngành: Sinh học thực nghiệm
Mã số: 60 42 30 LUẬN VĂN THẠC SĨ KHOA HỌC

NGƯỜI HƯỚNG DẪN KHOA HỌC: GS.TS LƯƠNG XUÂN HIẾN


2.2. 



 23
2.2.1. 



 23
2.2.2.  24
 24
 24
 25
 25
  29
 30
2.2.2.7. 





 Chlamydia trachomatis  ,




 
 31

33
 35
 37
 40
 43
 44
3.4.  44
KẾT LUẬN 49
TÀI LIỆU THAM KHẢO 50

58
BẢNG CHỮ VIẾT TẮT

C. trachomatis
Chlamydia trachomatis
DFA
Direct Fluorescent Antibody
EIAs
Enzyme Immuno Assays
LPS
Lipopolysaccharide
MOMP
Major Outer Membrane Protein
NAATs
Nucleic Acid Amplication Tests
PCR
Polymerase Chain Reaction
TPHA
Treponema Pallidum Hemagglutination Assay
1
MỞ ĐẦU
 
                V
sinh Chlamydia trachomatis 
  Chlamydia trachomatis ,
 



 Polymerase Chain
Reaction PCR) 


2
.  Chlamydia trachomatis
.

 c Nghiên cư
́
u ư
́
ng
dụng kỹ thuật PCR trong chẩn đoán viêm âm đạo, cổ tử cung do Chlamydia
trachomatis tại Bệnh viện Đại học Y Thái Bình 
-              
.
- X

  t 



  




g.
4
 
 C.trachomatis.
C.trachomatis 5  hu
T -t A, 
 D, E, F, G, H, I, 

[2,9].
C.trachomatis -
[34]. C.trachomatis   .
-    (Elementary Body- EB)        
m
-   (Reticulate Body- RB): Sau kC.trachomatis
     m),  
  -3  
 



  exocytosis).
 -1000   


1.2.  C.trachomatis (theo Yvonne Pannekoek).

5

VC.trachomatis        

C.trachomatis 
 
- C.trachomatis  (Human papilloma virus)  
.
Ở nam giới
C.trachomatis 
 
. 

.
C.trachomatis 

C.trachomatis 
 C.trachomatis
trong  .
1.2. Các xét nghiệm chẩn đoán vi khuẩn Chlamydia trachomatis [32]
1.2.1. Nuôi cấy
 C.trachomatis  vi
 C.trachomatis 
k 
 cC229.
-  
nhau 
-  
        )         
 
0
C- 5% CO2i

7

8
1.2.3. Kỹ thuật kháng thể huỳnh quang trực tiếp (direct fluorescent antibody- DFA)


 MOMP.

       

        
 lam 
,  
C.trachomatisC.trachomatis 

C.trachomatis  

X

. 
             C.trachomatis 
,  
               
NAATs   [32].
1.2.4. Kỹ thuật khuếch đại nucleic acid (nucleic acid amplification tests– NAATs).
NAATs 

 NAATs 
, 
 (trong 1 
 (Polymerase Chain Reaction PCR) 
             C.trachomatis [21]. N

10   (EB) 
C.trachomatis , cho 
.
DNA C.trachomatis. C
PCR 2
 DNA C.trachomatis. Enzym Taq polymerase 
DNA e DNA ch

DNA  
dSau , 


 agarose 1%, 

e








 .
C.trachomatis 
 ), 
.

10


 ()
  
 l C.trachomatis 

 et al.
  
   C.trachomatis  
 et al.
-  16  20
 


C.trachomatis 

 (Smith et al.
  
  
                
C.trachomatis 

 m C.trachomatis  
3.5% (5/145).     C.trachomatis   .    

 DFAC.trachomatis . et
al. 
 PCR.    

12
 vi khu  
12.C.trachomatis .2%.

13
   C.trachomatis    ,      
     ra  15  24 trong  . Trong 
2004,    19   847 100.000.  
1.087  trong  20-24.     
    C.trachomatis cao,       8    
. Nunavut C.trachomatis   (2.520
)    [27].
nested multiplex 
HO1, HO3, Chlam5/Chlam3, KL1/ Gon 5/Gon 3
 C.trachomatis trong 
 [19].   3 
  
    
tham gia. 15-20              
       -
  
 (Sigma  
2 1. 
C   Eppendorf)
        C.trachomatis.      
1
    
agarose 2%,  PCR .2
mM   dNTP; 3 mM MgCl2; 0.5 pmol/l   HO1, HO3, Chlam5 (5'-
CATTATGT-CGGAGTCTG-AGC- ), Chlam3 (5'-GGATGACTCAAGGAATAGT-
CG- 3), 1 l Internal Amplification Control (IAC), 0.4 pmol/l  IAC (5'-
TGTTTGACAGCTTATCAT), 5 l .5  
ion  l /15s, 60/30s, 72/
.2 mM dNTP; 2 mM MgCl2; 0.5 pmol/l 

   
 
  
 [20], 
C.trachomatis cao 
 
 C.trachomatis.  

15


Achchhe L Patel [11]  
            C.trachomatis 
Roche Amplicor test, DFA Trong   (2003
C.trachomatis  30.0%. 
 Surat,   b PACE2 test) 
8.5%,    Ahmedabad,           
C.trachomatis   

  Azerbaijan 3.1%  Bangladesh 3.4%. 
 Manila 23.3%, Cebu, Philippines 37.0%  14.0%  Nicaragoa [11] .
 () [12] 
ELISA  C.trachomatis 
         C.trachomatis    



58  -
..8 
.  

 [15]

 C.trachomatis 

   C.trachomatis 
t.8 ca ;
 . [18].
 C.trachomatis 
ao 


 



17
   
agaro  


 C.trachomatis.
 [22],
 
C.trachomatis v
 
   mM Tris (pH 9,0),
50 mM KCl, 3.5 mM MgCl2, 5          -
TTGCCTTAACCCCACCATT--CGTCCTTCCTAAAAGAG-CTA-
1.25 U Taq 
 


Nam [29]. 

39. 
        N.gonorrhoea    
C.trachomatis 

 
C.trachomatis 2.5%.
 
              
-1996 [24].
     

N.gonorrhoeae
             
C.trachomatis  
.4% cho giang mai, 3..8% cho
C.trachomatis, 5.V.

19
  
-  h [3].
  l
danh 
 . 

  C.trachomatis     
C.trachomatis 
 

 
  N.gonorrhoea  C.trachomatis 
C.trachomatis/C.trachomatis .9%, 48.4%, 54.9%.
C.trachomatis 
C.trachomatis 
 
 
   .    C.trachomatis       
 
 [1]    
              
C.trachomatis.
NC.trachomatis 
. 

20 [31]
C.trachomatis 
C.trachomatis 
 C.trachomatis 

. 
C.trachomatis 



Trích đoạn Điê ̣n di, phân tích kết quả Nồng độ MgCl2 Thời gian và nhiệt độ gắn mồi

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