O'IZZ-I
!
2010
^
BOYTli
BAO CAO
SAK
PHAM
NGIHtN ClJlJ »i TM CiP Bd
T«nd^t^i:
NGHlfiN
CCti
tJMG
DUNG
Qtl
TRINH
sAiN
XUIT
KIt4NG
HUYIT
TH4NH
sen,
B^
LifiT,
ROTA
SlJ
DIING
KJfiM
B.INH
vic
XIN

BOYTE
—BoCaca—
BAO CAO SAN
PHAM
NGHIEN CUtJ
DE TAI CAP BO
TSn
de
tai:
NGHIEN
ciru ifNG
DVNG
QUI
TRINH
SAN
XUAT KHANG HUYET THANH
Sdl,
BAI LIET, ROTA
SLT
DVNG
KIEM
DINH VAC XIN
Chu nhiem de tai: PGS.TS.LI Thi
LuSn
Off quan
(Th
cbvc) chii tri dl tai:
Trung tam Nghien
ciru
San xuat Vac xin va Sinh pham Y te

quan chu tri de tai:
Cap quan
ly:
Ma
s^
dk
tki
(n^u
co):
Then
gian
thu-c
hi^n:
Tong kinh
phi thifc hi^n
de tai:
Trong
d6:
kinh
phi
SNKH :
Ngucin kh^c
(neu
c6):
PGS.TS.Le Thj Luan
Trung
tSm
Nghien
cihi
San xuat Vac xin va Sinh

dl
tai: PGS.TS.Le Thi Luan
3.
Co quan chu
tri
de tai:
Trung tam Nghien cuu San xudt
Vic
xin va Sinh
phAm Y
t^
4.
Cff
quan quan
ly
de tai: Bg Y
t^
5.
Thir ky detail
6. Pho chu
nhifm
de tai
ho|lc
ban chii
nhifm
de tai (neu co):
7. Danh sach nhirng ngiroi thirc
hifn
chinh:
Nguyen Dang Hien

huyet thanh soi
5. Tieu chuan ca sa huyet thanh
b^i
lift
6. Tieu chuan ca
so*
huyet thanh Rota
7. Bai bao da dang
8. Dao
t^o
Thac sy
I. Qui trinh san xuat huyet thanh
sdi
Nguyen
vft
lieu
Khang nguyen Soi tit chung Edmonston do vien Kitasato cung cap;
Tho trong
lugng
ttr 1,8 ~ 2,5 kg;
Te bao Veto do vien Kitasato-Nhat ban cung cip;
Moi trucmg hoa
ch4t:
MEM,
NaHCOs
7,5%, BS, PBS (-), khang sinh P-S, Hank, chit
bao quan vacxin, ta dugc tiem dap ling mien dich (Adjuvant): Ta dugc hoan chinh
(CFA)
va ta dugc khong hoan chinh(IFA)
Vat lieu : Bom kim tiem trgn ta dugc, tu im CO2, pipet Aid, pipetman, diu tip cac loai,

truac
khi
thii
nghiem <4 don vi; Khoe manh,
khong bj bat ky benh ngoai da nao; Can nang: 2,0kg den 2,5 kg; Tho dugc
each
ly tu 7-10
ngay
Qui
trinh
gay ddp
irng mien
dich
tren
tho
Phucmg phap vien Kitasato-Nhat ban (Khang nguyen khong tinh che)
Stt
1
2
3
4
Ngay
0
07
14
28
Ngay thuc hien
10-09-2008
17-09-2008
24-09-2008

2 ben,
moi ben
2,5
ml
Tiem
(4)
10
(ml) VRS khong c6
chat phu gia
Tiem vao duoi da
diii
2 ben,
moi ben
5
ml
1.
Qui trinh san xuat huyet thanh soi
Nguyen vat
tifu
Khang nguyen Soi tir chung Edmonston do vien Kitasato cung cip;
Tho trong lugng tir 1,8 ~ 2,5 kg;
Te bao Vero do vien Kitasato-Nhat ban
cimg
cip;
Moi trucmg hoa
chit:
MEM,
NaHCOj
7,5%, BS, PBS (-), khang sinh P-S, Hank, chit
bao quan vacxin, ta dugc tiem dap ling

nguyen:
10'°CClD50/0,5ml
Tieu chuin tho
sir
dung:
Khang
thi
soi truoc khi thu nghiem <4 don vi; Khoe manh,
khong bi bit ky benh ngoai da nao; Can nang: 2,0kg
din
2,5 kg; Tho dugc
each
ly tir 7-10
ngay
Qui
trinh
gay ddp
ting mien
dich tren tho
Phuong phap vien Kitasato-Nhat ban (Khang nguyen khong tinh
chi)
Stt
1
2
3
4
Ngay
0
07
14

2 ben,
m6i ben
2,5
ml
Tiem (3) 5 (ml) VRS khong co
chat phu gia
Tiem vao duoi da
diii
2 ben,
moi ben
2,5
ml
Tiem (4)
10 (ml)
VRS khong co
chat phu gia
Tiem vao duoi da ddi 2 ben,
moi
ben 5
ml
5
6
42
49
22-10-2008
29 - 10 - 2008
Lay mau M2
(Lay het mau)
Tiem (5)
10

Phuong phap cua CDC-Hoa ky (Khang nguyen tinh sach qua
duone)
Stt
1
2
3
4
5
6
7
Ngay
0
14
21
28
35
49
Ngay thuc hien
10-09-2008
24 - 09 - 2008
01-10-2008
08-10-2008
15-10-2008
29-10-2008
Lay mau5 ml
tinh
mach tai
(Kiem tra khang
thi)
Lay mau Mo

thii
49 ke tir ngay gay mien
djch
miii 1)
Tai ngay thir 49 liy toan bg mau tho vao chai det, bao quan
4°C
qua dem , tach huyet
thanh, ly tam 3000 vong / phut trong 20
phiit,
lay huyit thanh, sau do bao quan -
SO'C
cho
toi khi sir dung.
Xac dinh hieu gia huyit thanh bang phucmg phap trung hoa tren tl bao vero mgt lop.
Huyit thanh dugc bit hoat
56°C
trong 30 phut, sau do dugc pha loang bac 2 va dugc trung
hoa vol
100
dcm vj khang nguyen soi,
hin
dich trung hoa dugc gay nhiim tren te bao vero
1 lop, sau 2 tuin dgc
kit
qua. Hieu gia khang
thi
la nlng do huyit thanh pha loang nhit ma
tji
d6
tjo

10%,
2% FBS
- Trypsin Versen va mgt
s6
dung djch khac
May
ly tam
Phi&n
nhua day det 96 gilng
Tu im 37°C va
36°C
- Pipet aid va pipetman cac loai
- Hood vo trimg va cac dyng cu khac phyc vu cho nghien cim
Phifffngphdp nghien
cicu.
Sdn xuit khdng nguyen bai
lift:
Nhan virut bai liet typ I, hoac typ II hoic
typIII
tren te bao vero mgt lop
vciri
nong do 1
CCIDjo
trong 300 tl bao vcri moi truang gay
nhiim DMEM tai nhiet dg
37°C
trong 72-96 gia, gat
hin
djch virut khi huy hoai tl bao
>90%.

djch tgo
khdng
the
bai li^t
Khdng
nguySn:
virut sabin typ
1
hoac typ 2 hoac typ 3
Him
liryng khdng
nguyen:
10'°CCID 50/0,1ml
Tho sir
dyng:
tieu chuin nhu tho
sii
dung cho san xuat huyet thanh scri
Cich gay dip
irng
mien djch: Phucmg phap CDC-Hoa Ky
Stt
1
2
3
4
5
6
7
Ngay

Tiem trong da nhieu
vi
tri, 0,1 ml/1 vi tri
Tiem (2)
1
+
1
(ml)
(VRRT+ta
dugc FIA)
_
Tiem vao co
diii
2 ben, moi ben 2 vj tri, moi vi ta'
0,5
ml
Tiem (3)
1
+
1
(ml) (VRRT+ta dugc
FL\)
Tiem vao co
diii
2 ben,
mil
ben 2 vi tri,
mli
vi tri
0.5 ml

: ly tam 3000
vong/phiit.l 0
phut. Thu huyit thanh bao quan
6
-20°C.
Kiem tra
hi^u
gid:
khang huyet thanh bang phucmg phap trung hoa vi lugng tren tl bao
Hep2C
3.
Qui trinh san xuat huyet thanh rota
Nguyen
vat
U^u
Khang nguyen
rota
typ G1P8,
G4P6,
RRV
T^
bao than tho Rherus
MA
104
Moi trucmg DMEM, 199
Tho tieu chuin
Ta
dugc
hoan chinh (CFA) va ta dugc khong hoan chinh(IFA)
Trypsin Versen va mgt

gia, sau
do hap phu virut len te bao tai nhiet do
37*'C
trong
1
gia
Bo het moi
truong
gay nhiim, nia te bao
01
lan bang moi
iruong
DMEM am khong
CO
trypsin, sau do cho them moi truang co trypsin vai nong do
10-20[ig/ml.
Dat chai te bao
da gay nhiim virut tai nhiet do
37''C
trong
3
ngay.
Gat
h6n
dich virut, dong tan
3
ISn,
ly
tam 3.000 v6ng/phut/30
phiit

) 0,2 + 0,2 (ml) (VRRT+ta dugc FCA)
Tiem trone da nhiiu vi tri. 0.1 ml/1 vi tri
2
3
4
5
6
7
14
28
42
56
70
84
LayTnauJvll
Lay mau M2
Lay mau M3
Lay mau M4
Lay mau M5
Lay mau M6
(Lay het mau)
Tiem (2)
1
+
1
(ml) (VRRT+ta dugc FIA)
_
Tiem vao ca dui 2 ben, mli ben 2 vi tri, mli vi tri 0,5
ml
Tiem (3)

1
(ml) (VRRT+ta dugc
FIA) _
Tiem vao co
diii
2 ben. moi ben 2 vi tri, mli vi tri 0,5
ml
—____^
Thu huyet
tiianh:
ly tam 3000 v6ng/phut,10 phut. Thu huyet thanh bao quan a < -
20 C cho toi khi sii dung.
Kiem
tra
hi^u
gid: khang the trung hoa bang phucmg phap ELIZA
4.
Tieu chuan khang huyet thanh Soi
Tieu chi
dinh
gia
Vo trimg
Hi$u
gia khang
huydt
thanh
Tinh dac hieu
Tieu chuan Nhat Ban
Vo trimg
>4.000

hi^u
khang
huyet
thanh
Typ I
Typ
II
Typ III
Typ I
Typ II
Typ III
Tieu chuin Nhat Ban
Vo trimg
>16.000
>
16.000
>
16.000
Cheo typ 2,
3
: <32
Cheo typ 2, 3 : <32
Cheo typ 2, 3 : <32
Tieu chuan
ca
so*
Vo trimg
>65.000 cho thit
nghi?m
xac dinh tac

sa
Vo trimg
>
6.400
gia
khang
huyit
thanh
Tinh dac
hieu khang
huyet
thanh
Wa
DSl
RRV
RRVmix
G1P8
Wa
DSl
RRV
RRVmix
5.000
5.000
Dac hieu bang thir nghiem
nhan dang qua ky thuat
miin
dich huynh quang
> 1.000.000
>100.000
> 1.000.000

si
60.62.50
Ba bao ve vol diem xult
sac
nam 2008
Bp GIAO
DIIC
VA DAO TAO
TRUOfNG
DAI HOC NONG NGHIEP HA NOI
ca
NGUYEN THI NGA
NGHIEN
Ciru
SAN
XUAx
KHANG THE BAI LIET TREN THO
VA DANH GIA TINH
AN
TOAN
CUA VAc XIN BAI LIET
TRONG PHONG THI NGHIEM
LUAN VAN THAC
SI
NONG NGHIEP
Chuyen nganh : THU V
Mas3
: 60.62.50
Nguoi huong dan khoa hoc: PGS.TS LE THI LUAN
HA

sir
phan bo 5
2.3.
0 Viet
Nam
5
2.4. Dac diem virus bai liet 5
2.5.
Benli
bai liet a ngiroi
13
2.6.
Tieu chuan Viet Nam ve
phuonng
phap kiem tra tinh an toan
ctia
vacxin phong bai liet
16
2.7.
Vacxin phong benh bai liet
18
2.8.
Nghien cuu san xuat khang the bai liet 21
3.
DOI TUONG,
DIA
DIEM,
NOl
DUNG VA PHUONG PHAP
NGHIEN

Ket qua san
xuk
khang the tir huyet thanh Tho
\kn
thir nhat
51
4.1.3.
Kit
qua san xuat khang
thS
tir huyet thanh Tho lan thir hai 57
4 2.
Kit
qua kiem tra tinh an toan vacxin bai liet trong phong thi
nghiem 63
^•2.1.
Kiem tra nguyen lieu dau cho san xuat vacxin bai Het - khi san
xuat 63
Ket qua kiem tra te bao sir dung cho san xuat vacxin bai
Met
trong
bay nam tit nam 2000 - 2006 65
4
2.3.
Kit
qua kiem tra an toan vacxin ban thanh pham
4-3.
Ban luan
4.3.1.
Nguyen heu