


transcription polymerase chain reaction







Abstract. T       

-P
-
       -


Keywords. ; ; 

Content
I. Đặt vấn đề

Orthomyxoviridae. 















, 






vir

/H5N1  . 

1. a chn p  thc hin phn ng multiplex RT-

2. n cu ti n ca phn ng multiplex RT-PCR: nhit
 i gian gn mi, nng  
3. Th nghin ng multiplex RT-t s
mu bnh phm thu thm.
II. Vật liệu và phương pháp nghiên cứu
Mâ
̃
u bệnh phẩm:  




 g); 30

Tách chiết RNA: thu RNA  

Thiết kế mô
̀
i :  5, N1 
, 2.0.11. Sau khi


 5.4 /H5N1 




















 



 1000 (Biorad). 

(10 ) 




2%, 



 ethidium bromide 0,5 /ml 15 , 


















A/H1N1, /H3,  (







 ), /H5
(





 ) 

















/H5N1 1.2/blunt (Fermentas) , 















Kit (Fermentas). 


















. Sau


, 















 10

.
III. Kết qu
Thiê
́
t kê
́
mô
̀
i: 







 5, N1 




 , 3 









DiagMR
GTGACAGGATTGGTCTTGTCTT
158-139
55.8
DiagH5F
AGTGATCAGATTTGCATTGGTTAC
46-69
54.6
371 bp
DiagH5R
GACCAAGAACTTTTGGGGATG
416-396
55.4
DiagN1F
CCAGTTGGTTGACAATTGGAAT
503-524
54.5
292 bp
DiagN1R
GCATCAGGATAACAGGAGCA
794-775
55.4
Tối ưu phn ứng mutiplex RT-PCR: -












) . 

, 
mutiplex RT-









3 














95
o
C trong 15 ; 

45 

(94
o
C: 30 ; 57
o
C: 30 ; 72
o
C:
30 ); 



































 (ngươ
̀
i, Escherichia coli, Mycobacteria tuberculosis,
Chlamydia trachomatis, Neisseria gorrnorhea, Hepatitis B virus, Hepatitis C virus,
Human immunodeficiency virus, Human Papillomavirus),  utiplex RT-PCR
.

Ảnh điện di đa
́
nh gia
́

n ư
́
ng mutiplex RT-PCR:
 , 

10
1
10
10

/





 10 



 2%. 



 di cho






nha
̣
y cu
̉
a pha
̉
n ư
́
ng multiplex RT-PCR
M: thang DNA 100 bp; NC: Chư
́
ng âm; S lưng bản sao RNA trong mu th đưc ghi
ch  pha trên.
ng dng k thuật mutiplex RT-PCR pha
́
t hiê
̣
n cu
́
m A /H5N1 trên mâ
̃
u bê
̣
nh
phâ
̉
m:
Kmutiplex RT-PCR 







). 






utiplex RT-









14 





.

Kết qua

m pha
̉
n ư
́
ng multiplex RT-PCR phát hiện virus cm gia cầm A/H5N1
trên một s mu bê
̣
nh phâ
̉
m
M: Thang DNA chuẩn 100 bp; NC: Chứng âm; PC: Chứng dương;
1-10: Mu bệnh phẩm.

Kết luận
- 
mutiplex RT-PCR.
- 









 -














/H5N1 .
- ex RT-PCR 

References :
Tiếng Việt
 Báo cáo tình hình cm gia cầm tại Việt Nam, 



Tạp ch Công nghệ Sinh học 2(1): 1-18.
Tạp ch Khoa học Kỹ thuật Th y, 11(1): 81-86.
Tạp ch Khoa học Kỹ thuật Th y,
11(2): 53-58.
Tiếng Anh
5. Abdel-Ghafar AN, Chotpitayasunondh T, Gao Z, Hayden FG, Nguyen DH, et al,
N. Engl.
J. Med, 358, pp. 261-73.
6. Apisarnthanarak A, Kitphati R, Thongphubeth K, Patoomanunt P, et al, 2004,
Emerg Infect Dis, 10, pp. 1321- 4.
-length

            
influenza A (H5N1) in a child presenting with diarrhea foll N
Engl J Med, 352, pp. 686-91.
19. De Jong MD, Simmons CP, Thanh TT, Hien VM, Smith GJ, Chau TN, Hoang DM,
Chau NV, Khanh TH, Dong VC, Qui PT, Cam BV, Ha DQ, Guan Y, Peiris JS,
           uenza A
Nat Med, 12,
pp. 1203- 07.
20. De Jong MD, Tran TT, Truong HK, Vo MH, Smith GJ, Nguyen VC, Bach VC, Phan
e
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