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8122
2010
BdYTf
BAO
CAO
idl QUA NGElfiN Ctfl] fli
T.4l
Clp
BO
nUESM
ecu
rjNG DtJNG
QUI
TRINH
SAN
XUXT
KHANG
HIJYf
F
THANH
Sdl,
BAILE^,
ROTA StJTOWG SQ^M BBStH vAC SIN
Cbunhttind^ai:
PGS.TS.L«TMl«te
Cff quan (Td ch6c) did
hi
S«
Oil:
Trung tftm Ngbi«n c«iu
Sin

%.u&
Vic
siii
v^
Sinfe phlas
Y
t*
Nam
- 2010
BOYTE
—SOQGS—
BAO CAO
KET
QUA NGHIEN
ClTU
DE
TAI
CAP
BO
Ten
de
tai:
NGHIEN
Ctru tTNG DVNG
QUI
TRINH
SAN
XUAT
KHANG
HUYET

THANH SOU, BAI
LIfT,
ROTA
SlJ DVNG KIEM SpjH
VAC
XIN
Chu nhifm
de
tii:
PGS.TS.Lg
Thi Luan
Ctf
quan chu
tri
de tai: Trung
tam
Nghien
cuu
San xuat
V^c
xin
va
Sinh
pham
Y te
Cap quan
ly:
B9
Y te
Ma so de tai

(neu
c6):
Tlioi
gian
tliirc
hi^n:
ttr thang 7 nam 2007 den thang
12
nam 2009
Tong
Idnh plii tliuc liif
n de tai: 910
trif
u
dong
TrODg do: Idnh phi SNKH
:
598
trifu
dong
Nguon khac (neu co);
Nam 2010
p(/0
(
2MO
Boo
cdo
kit qua nghien
cuu
Se tai cap Bo

1.
Ten de tai:
NGHIEN
ctru tTNG DVNG
QUI TRINH
SAN
XUAT
KHANG HUYET
THANH
sen,
BAI
LIET,
ROTA
SU"
DVNG
KIEM
DpVH
VAC XIN
2.
Chu
nhifm
de tai: PGS.TS.Le
Thj
Luan
3.
Cor
quan chu tri de tai:
Trung tam Nghien ciru, San xuat Vac xin va Sinh pham Y te
4.
Cv quan quan ly de tai:

B9
Y te
5.
Thu-
Ity
de tai:
6. Pho chu
nhif
m de tai
ho^c
ban chii
nhi|m
de tai (neu c6):
7. Danh sach
nhimg nguM thuc
hien chinh:
Nguyin DangHiSn
Nguyen
Thj
Quy
Le
Trung Diing
Nguyin
Thiiy
Huong
Nguyen Thi Mai Huong
TrSn
Bich Hanh
Dang Mai Dung
Nguyin

NO
Anh Thu
Ngo Thu Huong
8. Cac
dl
tai
nhanh
(de
mi^c)
ciia de tai (neu c6)
9.
ThM
gian
th^c hi|n
de tai: tu thang 7 nam 2007
dSn
thang
12
nam 2009
Cac chir viet tat
BS (Bovin serum)
PBS
(Photphat
buffer solution)
P-S (Penixilin - Streptomycin)
MEM (Minimum Eagle Medium)
FBS (Fetal Bovine Serum)
: Huyet
thanh
be

: Dung
dich
dem
muoi
phot phat
: Khang sinh Penixilin va Streptomycin
: Moi
truong
Eagle toi thieu
: Huyet thanh be bao thai
DMEM (Dubeco Minimum Essencial Medium): Moi truong
ca
ban
t6i thi6u
CPE (Cytopathic effects)
:
Huy hoai
tk
bao
FFU(fluorescence
forcus unit) : Bon vi mien dich huynh quang
CCID50(cell culture infectious dose 50)
:
Lieu
gdy nhilm
50%
tl
bao
IgG
(Immuno Globulin

0)
IgA
(Immuno Globulin A)
IgM
(Immuno Globulin M)
IgE
(Immuno Globulin E)
IgD
(Immuno Globulin D)
MAI
04
Vero
Hep2
Da
FIA
(Freund's Incomplete Adjuvan)
FCA (Freund's Complete Adjuvan)
WHO (World Health Organization)
: Khang the IgG
: Khang
thi
IgA
: Khang the IgA
: Khang
thi
IgA
: Khang the IgA
: Te bao than bao thai
khi
Rhesus

:
T8 bslo than
khi xanh Chau Phi
; Te bao ung thu thanh quan
nguai
: Dalton
: Ta
dirge
khong hoan toan
: Ta dugc hoan toan
: To
chuc
Y te The
gi(5i
Bao cao torn tat de tai
Bit
van
d€
Trung tam Nghien
ciiu
San
xult
Vic xin vi Sinh
phSm
Y te hay Trung tam
NghiSn
cuu San xult Vac xin Sabin
dugc
thanh lap
tii nam

1994 c6 nhiem vu san xuat
v4 kiem dinh
vac xin. Hien nay Trung tam dang san xuat va kiem
dinh
vac xin Bai
liet,
vac xin Soi; nghien cuu, san xuat vac xin Rota phong
tieu
chay cho tre em. He thong
kiem dinh cua Trung Tam duoc dao tao co ban va chuyen sau phuc vu cong tac kiem
djnh
chat
lucjmg
vac xin. Mot trong nhung nguyen vat lieu co ban sit dung danh gia an
to4n
cua vac xin bin thanh pham trong phong thi nghiem la
huySt
thanh khang Soi,
khang virut Bai lift,
vk
khang virut Rota. Khang thi nay
s\jt
dung cho
thii
nghiem tim
tdc
nhan ngoai
lai
trong h5n djch vac xin san xuat.
D5ng

thoi khang the nay ciing
sii
dung de chuan
dO
hieu gia
vie
xin Rota va
vie
xin Bai liet thanh
phAm.
Nhiing san
pham nay
truac dJy
dugc cung cap hoac mua tir cac nha san xuat Nhat, My. Tuy nhien
de chu
dgng
trong cong
vi?c,
Trung tam thuc hien de tai "Nghien
ciiu ling
dung quy
trinh san
xudt khdng huySt
thanh Soi, Bai
liet.
Rota
sii
dung
kiSm
dinh

vie
xin"
vdi
nhiing muc dich sau:
1.
LTng dijng
vk hoan thien qui trinh san xuat
huySt
thanh dac
hiSu vurut
Soi,
Bai
lift.
Rota
sii dymg
trong
kifm
djnh vac xin;
2.
San xuit dugc
huySt
thanh khang vurut Soi. Bai
liet.
Rota dat tieu chuin co
sd;
3.
Danh gia chat
lugng
huyet thanh ve tinh dac hieu, hieu gia so vdi
huySt

thanh nhap ngoai
Nguyen
v|t
lien
va
phinrag
phap
tiln
hanh
De
CO
dugc khang the,
trade
het de tai
pliai
san xuat khang nguyen, phuong
phap san xuat khang nguyen Soi, Bai
liet.
Rota la
sii
dung chung virut da dugc cung
cap tir cac phong thi nghiem chuan quoc
tS hojc
chung san xuat vac xin nhan
len
tren
nuoi cay te bao thich hop
vdi
the tich va hieu gia du de sau khi tinh sach gay dap ling
miln

djch t6t.
Sau khi nhan
virat
tren te bao, h5n dich virut dugc dong tan, ly tam, lay
nude
noi sau do tinh sach qua dudng Sacahrose
40%,
lay can, hoa can , ly tam 15.000
vdng
/phut/40
phiit,
lay
nude
noi, dieu chinh sao cho hieu gia theo lieu gay nhilm thich hgp
ciia
timg
loji
virut:
10*'° CCID
50/0,5ml cho virat Soi;
10''° CCID
50/0,lml cho virat
B?ii
liet va
lO''"
ffii/lml
cho virat Rota.
Thd
dugc mua tii trai thd Ba vi trgng lugng tir 1,5 kg
din

2,5 kg, khoe manh,
dugc
cich
ly it nhat 10 ngay
trade
khi
sii
d\mg,
khang
thi
khong qua 4 don vi vdi
khang
nguygn die
hieu. Don
vj
khang
thi
d
day
dugc hieu la
hi?u
gia cua khang
huylt
thanh vi
dij:
Hifu
gia khang huyet thanh khi chuin
df
cho ket qua 4.000 va dugc coi la
4.000 don

vi
khang huyet thanh.
Sii
d\mg
ta
chit hoan chinh (FCA) hoac ta chit khong hoan chinh (FIA) pha
chl
h5n dich khang
nguySn,
tiiy loai
khang nguyen c6
Hch v4 lilu
ti6m khac nhau.
Doi vdi virat Sdi cd 2 phuong phap tao khang
thi,
do la phuong phap cua vien
Kitasato
Nh|t
Ban va phuong phap cua Trung tam phong
chdng va
kiem soat
b$nh
tat
CDC-Hoa
k^.
hai phuong phap
niy
khac nhau ve chuin bi khang nguyen,
va lich
tiem

cv thi
nhu sau:
Phuong phap san xuat khang huylt thanh Sdi khong qua tinh sach vk phuong
phap gay
dap
ling miln dich
trSn
tho (Phuong phap vien Kitasato-Nhat ban)
Thuc
hifn
5
lin
gay dap ling miln dich theo lich ngay: 0 : 7 : 14: 28 : 42. Lay
mau (5ml) trade
khi tiem lin l(Mo), 5ml trudc khi gay mien dich
mQi
4 (Ml)va
liy
toto b$ mau vko
ngay
thii
49 sau tiem
Ian
1(M2), nhu
vjy
se co 3 mau huylt thanh
kiem tra
hif
u gia.
Lilu lugng tiem moi

Ian
nhu sau:
Tiem lin
1
ngay
0:
Tiem 2,5ml
hdn
dich virut
+
2,5
ml ta chit
FCA;
Tiem trong
da
nhilu
vj tri,
0,1
ml/1
vi
tri;
Tiem
Ian
2 ngay
7:
Tiem 5 ml virat Sdi khong cd ta chat; Tiem vao dudi da
diii
2
bta,
m6i bSn

2,5 ml;
Tiem lin
3
ngay
14:
Tiem
5
ml virat Sdi khong cd ta chat; Tiem
vao
dudi da
dCii
2
bgn,
m5i ben 2,5 ml;
Tiem
Ian 4 ngky
28:
Tiem
10
ml virut Sdi khong cd ta chat; Tiem vao dudi da
diii
2 ben,
mdi
ben
5
ml;
Tiem
Ian 5
ngay
42:

Tiem 10 ml virat Sdi khong cd ta chat; Tiem vao
tinh
mach tai
2
ben, m5i ben
5
ml.
Phnvng
phap san xuat khang huyet thanh Bai
liet.
Rota theo phuong phap gay
d4p
ung miln djch cua Trung tam Phong
chong va
kiem soat benh tat CDC-Hoa
Ky
Thd dugc
each
ly tir 7-10 ngay, gay
dap
img mien dich 6
Ian
tiem
each
nhau 2
tuin
vko
ngay:
0:14:28:42:56:70;
84 liy mau tai 5ml

trade
khi tiem lin
1 va
ngay
trade
khi tiem
cao Ian
sau, lay
mau
toan
bg
cu6i
cilng vao
ngay 84 sau lin
tifm
1.
Nhu vay se
cd 7
mlu
huylt thanh
(MO,
Ml,
M2,
M3,
M4,
M5,
M6)
hoac se co 6 miu
mau
khi Hch

liy
mau
0:28:42:56:70:84 (Mo, Ml,
M2,
M3,
M4,
M5)
Lich
va
lilu tiem cu
thi:
Tiem lin
1
ngay
0:
Tiem
0,2nil
virat
B^i
li?t +
0,2ml
ta dugc FCA; Tiem trong
da
nhilu
vi
tri, 0,1 ml/1 vi tri;
Tiem
Ian
2 ngay
14:

Tiem
1ml
virat Bai liet
+
1ml ta dugc FCA; Tiem vao co
dii
2
bfn,
mSi
ben 0,5 ml;
Tiem
Ian 3 ngay
28:
Tiem
1ml
virat
B?i
lift
+
1ml ta dugc FCA;
TiSm
vao co
diii
2 ben,
mdi bto
0,5 ml;
Tiem lin
4
ngay
42:

Tiem
1ml
virat
Bai
lift
+
1ml ta dugc FCA; Tiem vao co
diii
2 ben,
mdi bfn
0,5 ml;
Tiem lin
5
ngay
56:
Tiem 1ml virat
Bai
lift
+
1ml ta dugc FCA; Tiem vao co
dCii
2 ben,
mSi
ben 0,5 ml;
Tiem
ISn
6 ngay
70:
Tiem 1ml virat Bai lift
+

1ml ta dugc FCA; Tiem vao
eg
diii
2 ben,
mdi
ben 0,5 ml;
Sau mSi lin tiem, lay
mau kilm
tra diin bien
dap
ling khang the
va
chon ra qui
trinh thich hgp cho hieu gia khang the cao nhat.
Sii
dung phuong phap trang hoa vi lugng tren te bao vero xac dinh khang huylt
thanh Sdi
va
phuong phap tnmg hoa vi lugng tren te bao Hep2
xac
dinh khang huylt
thanh Bai lift. Xac dinh khang huylt thanh Rota
bang
phuong phap
ELISA.
Dong
thdi
sii dung phuong phap trung hoa de ling dung khang the trong
thii
nghif

m tim tac nhan ngoai lai trong
hdn
dich virat vacxin khi trang hoa hoan toan me
gjt
don. Tim ra lugng khang huylt thanh trang hoa hoan toan me gat don va ap ung
trong kiem dinh
vie
xin;
Khang huyet thanh san xuat ra con dugc
sii dmg
vao thu nghiem xac dinh nhan
dang virut vacxin va xac dinh hieu gia virat vacxin bang each tim don vi khang
thi ap
dung cho thu nghiem nay.
Ket qua
1.
Qui
trinh
san
xu^t
khang huylt thanh
l.I.
Qui trinh san xuat khang huyet thanh Soi
Bu&c
1. San
xudt
khang nguyen
S&i
Viras Edmonston Vero7 dugc san xuat tren te bao vero tao virat dugc ggi la
Edmonston Vero 7/P2.

H5n
dich virat nay sau khi gat dugc ly tam bd te bao va sau do
dugc
Igc
vdi mang
Igc
0,65.
Hdn
dich dugc
thii v6 triing
va kiem tra hieu gia, hifu gia
SU
dung cho gay
dap ling
miln dich la
10'-°CCID
50/0,5ml
Buac
2. Chon
Igc
va
each
ly tho. Thd mua
vl
dugc
each
ly trong 7-10 ngay, thd
dugc kiem tra
v6 can
njng,

benh ngoai da, huylt thanh Sdi. Tieu chuan tho
sii
dung:
Khang huyet thanh Sdi
trade klii thii
nghifm <4 don vi; Khde manh, khong
M
bit ky
bfnh ngoai da nao; Can
njng:
2,0kg
din
2,5 kg; Thd dugc
each
ly tir 7-10 ngay. Qui
trinh gay dap img mien dich tao
ichdng
huyet thanh S&i tren tho,
/"huong
phap vifn
Kitasato-Nh$t
ban
Stt
1
2
3
4
5
6
NgJy

0
07
14
28
42
49
Lay mau
Mo
Ml
Lay mau
gitia
ky
M2
Lay
mau
toan
bo
Cach
tiSm
Tiem 2,5ml hdn dich virat + 2,5 ml ta chit FCA;
TiSm
trong da nhilu vi tri, 0,1 ml/1 vi tri
Tifm 5 ml virat Sdi khong cd ta chat; Tiem
vao
dudi da
diii
2 ben, moi ben 2,5 ml
Tiem 5 ml virat Sdi khong cd ta chit; Tiem vao
dudi da
diii

2
bSn,
moi
b6n
2,5 ml
Tiem 10 ml virat Sdi khong cd ta chat; Tiem vao
dudi da
diii
2 ben, moi ben 5 ml;
Tiem 10 ml virut Sdi khong co ta chat; Tiem
vao
tinh
mach tai 2 ben, moi ben 5 ml
Budc3.Lay
mau
Lay
mau
kiem tra huyet thanh Sdi 3
Ian,
lin 1
trade
khi gay miln dich
mili
1;
Lan 2 lay mau tai
trade
khi gay mien dich miii 4 de kiem tra kha nang tao huyet thanh
giila
ky; Lan 3 lay mau toan bg sau 7 ngay gay miln dich miii 5 (ngay
thii

49 ke tit
ngay
gay
miln dich miii 1)
Tai ngay thii 49 liy toan bg mau thd vao chai det, bao quan
4*'C
qua
dSm
, tach
huylt thanh, ly tam 3000 vong /
phiit
trong 20
phiit,
liy huylt thanh, sau dd bao quan -
30°C cho tdi khi
sii
dung.
Budc 4. Kilm dinh huylt thanh
Kif
m tra v6 khuin
xac
dinh hieu gia
huyft
thanh
bjng
phuong phap trung hoa tren te bao vero
mdt
Idp.
Huyet thanh dugc bat boat
56''C

trong 30 phiit, sau do dugc pha
loang
bac 2 va
dugc trang
hda
vdi 100 dgn vi khang nguyen Sdi, hon dich trung hoa dugc gay nhiim
tren te bao vero 1 lop, sau 2 tuin doc ket qua. Hifu gia khang huylt thanh la nong do
huyet thanh pha loang nhat ma tai dd tao ra phan ling trang hoa
(tl
bao binh thudng
khdng bi huy hoai)
1.2.
Qui trinh san xuat khdng huylt thanh Bai lift
Bird^c
1.
S&n
xuat
kh^ng
nguyen
San xudt
Ididng nguyen
bgi
lift:
Nhan virut bai
lift
typ
1,
ho5c
typ II
hoSc typlll tr6n

tl
bao
vero
mpt Idp
vdi nong
d0 1 CCID50
trong 300 te bao vdi moi
trucrag
gay nhiem DMEM
t^ii
nhift d0 37°C
trong 72-96 gid,
gSt
hon dich virut khi huy hoai tl bao >90%.
Tinh sgch
khdng
nguyen:
HSn dich virut sau khi dugc dong tan 3 lan, ly tam 3000
vdng/phut
trong 30 phut,
Idy nude
noi. Can dugc sieu am va ly tam lay
nude
noi.
Nude
no:
hpn
lai va dugc
loc
vdi co

Igc
lan
lugt
0,65
pm,
0,45
^m,
0,2
|xm.
Hon dich virut dugc sieu
Ipc
vdi c8 dac
100-200
lin va dugc sieu
li
tam 36.000
v6ng/phuf3
gid tai
4''c,
lay can, hoa
loang
c^
bing EMS, pha loang sao cho nong do virut
lO''
CCED50/0,lml.
Hon dich virut
niy
dugc chia nho va bao quan tai
-80°C
cho tdi khi

sii
dung.
Chi dong tan
01
lin trudc khi
sii
d\ing.
Budc
2.
Chpn loc
vd cdch ly tho. Tho mua ve dugc
each
ly trong
7-10
ngay, tho dugc kiem
tra vl
cSn
n^g,
bfnh ngoai
da,
huyet thanh Sdi. Tieu chuan tho sic dung: Khdng huyit
thanh
B?i Uft trade
khi thu nghiem <4 don vi; Khoe manh, khong bi bat ky benh ngoai da
nao;
can
n^g:
2,0kg
din
2,5 kg; Thd dugc each ly tii 7-10 ngay. Qui trinh gay dap img

miin dich tao khdng huyit thanh Bai liet tren tho- Qui trinh Trung
torn
Phong chong vd
Kiim sodt
benh
tat
CDC-Hoa
Ky
Stt
1
2
3
4
5
6
7
Ngay
0
14
28
42
56
70
84
Lay
mau
Mo
Lay
mau tai
Ml

Lay
mau
tai
M2
Liy
mau
tai
M3
Lay man tai
M4
Liy
mau
tai
M5
Liy
mau
tai
M6
(Liy
mau
dgng
mach
catih)
each
tiem
Tiem 0,2ml virat Bai
hft
+
0,2nil
ta dugc

FCA;
TiSm
trong da
nliieu
vi tri, 0,1 ml/1
vi
tri;
Tiem 1ml virat Bai lift + 1ml ta dugc FIA;
Tiem
vao
co dui 2
bSn,
m§i ben 0,5 ml;
Tiem 1ml virat Bai lift + 1ml ta dugc FIA;
Tiem vao co
diii
2
bSn,
mSi ben 0,5 ml;
Tiem 1ml virat Bai lift + 1ml ta dugc FIA;
Tiem
vao
co
diii
2 ben,
mdi
ben 0,5 ml;
Tiem 1ml vurut Bai lift + 1ml ta dugc FIA;
Tiem vko co dui 2 ben, mdi ben 0,5 ml;
Tiem 1ml virat Bai lift + 1ml ta dugc FIA;

Tifm vko co
dtii
2 bfn, moi
bSn
0,5 ml;
Bu&c
3. Lay mau vd thu huyet thanh
Liy
mau
7 lin, lin
thii
nhit trade khi gay
dap ling
miln dich lan 1,
cac
lan tiep
theo liy
mku
ngay trade mdi lin gay dap irng mien dich tiep theo; lan cudi cung lay
toan bg
mau vao
ngay
thii
84.
Mau
dugc ly
tSm
3.000
vdng/phut/10
phiit, liy huyet thanh, bao quan huylt

thanh ri6ng
rg
cua timg thd tai nhift
dp -20°C
cho tdi khi su
diing
Bit&c
4. Kiim dinh huyet thanh
Kiem tra v6 khuin
X4c
dinh hifu gia huylt thanh bing phuong phap trung
hoa
trfn tl bao Hep2
mOt
Idp. Huylt thanh dugc bit boat 56°C trong 30 phiit, sau dd dugc pha loang bac 2
va dugc trung hda vdi 100 don vi khang nguyen Bai lift, hdn dich trung hoa dugc
gjy
nhiim tren te
bao
Hep2, sau 5-7 ngay doc ket qua. Hifu gia khang huyet thanh la nong
dg huyet thanh pha loang nhit ma tai dd tao ra phan ling trang hoa (te bao binh thudng
khdng bi buy hoai)
1.3. Qui trinh san xuat khang
hnyet
thanh Rota
Budc
1.
Son xudt
khdng
nguyen

TI
bao
MA104 mot lap hoac tl bao vero mot
Idp,
dep dugc
sii
dung cho su nhan len
ciia virat Rota. Te bao dugc nia 2 lan vdi mdi truong DMEM dm, sau do dugc dat tai
37'*C
it
nhat
3
gid.
Virat dugc hoat hoa vdi trypsin nong dp
15nl/ml
tai nhift dd
37°C
trong
1
gid, sau do
hip phu
vmit
len tl bao tai nhift dp 37°C trong
1
gid
Bo
hgt
moi tradng gay nhiem, rua te bao
01
lan bing moi

tiudng
DMEM am khong cd
trypsin, sau do cho them moi tradng co trypsin vdi ndng dp
10-20^g/ml.
Dat chai te bao da
gay
nhiem virat tai nhift dp
37°C
trong
3
ngay.
Gat hon dich virat, dong tan
3
lan, ly tam 3.000 vong/phut/30 phut bd can.
Nude nil
thu dugc sieu ly tam 25.000 v6ng/phut/2 gid tai
4°C
vdi nong dp dudng
40%.
Bd
nude
ndi,
thu
c^
va hda
c5n
vdi TNC, sau do pha loang khang nguyen thu dugc sau
cho lugng virat khoang
lO'-lO*
fiii/ml

Buac
2.
Chpn
IQC
vd cdch ly tho
Thd mua vl dugc
each
ly trong 7-10 ngay, thd dugc kiem tra
vS cSn
nang, benh ngoai
da, huylt thanh Sdi. Tieu chuin tho sir
dung:
Khdng huyit thanh Rota
trade
khi thir nghiem
<4 don vi; Khde
mjnh,
khdng bi bit ky bfnh ngoai da nao; Can nang: 2,0kg
din
2,5 kg; Tho
dugc
each
ly
tir
7-10 ngay. Qui trinh gay dap img
mien
dich tao khdng huyet
thanh
Rota tren
thd

-Qui
trinh Trung tdm Phong chSng va Kiem sodt benh tat CDC-Hoa
Ky(gi6ng
nhu qui
trinh
tiem sdn
xudt
khdng
huyet
thanh
Bai Hit)
Budc 3. Liy
mau va
thu huylt thanh(gi6ng nhu
san
xuit khang huylt thanh Bai
lift)
Budc 4. Kilm dinh khang huyet thanh
Kilm tra vd khuin
xac
dinh hifu gia khang huylt thanh bang phuong phap ELISA. huyet thanh
dugc pha loang bac 4 .
Hifu
gia huylt thanh
la
ndng do pha loang nhat
ma
tai dd xay
ra
phdn

img khang nguyen khang
thi.
2.
Tieu chuan cor
sd
2.1.Tieu chuan
khing
huyet thanh Sdi
Tieu chi danh gia
Vdtrung
Hifu
gia khang huylt
thanh
Tinh dac hieu
Tien chuan Nhat Ban
Vdtrung
>4.000
Die
hieu bang
thii
nghiem
nhan dang qua ky thuat
mien dich huynh quang
Tieu chuan co
sd
Vdtrung
>4.000 cho
thii
nghiem xac dinh
tac

nh4n
ngoai lai
>1.000
cho
thii
nghifm nhan dang
virat Sdi
Dac hifu bang
thii
nghiem nhan
dang qua ky thuat miln dich huynh
quang
2.2.TiSu
chuan
khing huySt
thanh Bai liet
Tieu chi danh gia
Vdtrung
Hifu
gia
khang
huyet
thanh
bai
lift
Tinh
djc
hifu
khang
huyet

thanh
Typ
I
Typ
II
Typ
III
Typ
I
Typ
II
Typ
III
Tieu chuan Nhat Ban
Vdtrung
>
16.000
>16.000
>16.000
Cheo typ 2, 3 : <32
Cheo typ 2, 3 : <32
Cheo typ 2, 3 : <32
Tieu chuan co sd
Vdtrung
>65.000 cho
thii
nghiem
xac
dinh
tac nhan ngoai lai

<65.000 cho thii nghiem xac dinh
hifu
gia vie xin thanh phim
>8.000 cho thu nghifm xac dinh
tac
nhan ngoai lai
<8.000 cho
thii
nghiem xac dinh
hifu
gia
vie
xin thanh phim
>16.000 cho thir nghifm
xac
dinh
tac nhan ngoai lai
<16.000 cho thu nghifm
xac
dinh
hifu
gia vac xin thanh phim
Cheo typ 2, 3 ; <32
Cheo vdi
typ 1,3: <16
Cheo vdi typ
1,2
: <32
2.3.Tieu
chuan

khing
huyet thanh Rota
Tieu
cM
danh gia
Vdtrung
Hifu
gia
khang
huyet
thanh
Tinh dac
hifu
khdng
huyet
thanh
G1P8
Wa
DSl
RRV
RRVmix
G1P8
Wa
DSl
RRV
RRVmix
Tieu chuin Hoa Ky
Vdtrung
5.000
5.000

Dac hieu bing
thir nghifm
nhan dang qua
ky
thuat
miln dich huynh quang
Tif
u chuan co sd
Vdtrung
> 6.400
>
1.000.000
>100.000
>
1.000.000
> 100.000
Dac hieu bang thir nghiem
nhan dang qua
ky
thuat miln
dich huynh quang
3.
So
Iirgng va
chat
lu-gng
huylt
thanh
san xuat
a.l.Khing

Huyet thanh Sdi
Dgt
san
xuat
1
Tong 1
Sdthd
1
2
3
4
5
6
7
8
9
10
10
tho
Huyft thanh > 4.000
Dat
tif
u chuin
Nhjt
Ban
Thi
tich
43
42,5
23,0

25,5
134
Hifu
gia
4.096
8.192
4.096
4.096
Huyet thanh < 4.000
Dat TC Co sd
The tich
35
42,0
51,0
128
Hieu
gia
3.072
1.536
1.536
Khdng dat
Thf tich
20,0
46,5
38,5
105
Hifu gia
96,0
256
512

2
Tong
2
3
Tong
3
Tong
cgng
13
19
25
26
27
28
29
30
31
33
10 thd
34
35
36
37
38
39
40
41
42
9 thd
29 thd

38
42
80
45
45
259
(22,78%)
6.144
8.192
6.144
41
35
37
45
41
35
40
274
30
25
50
46
45
42
40
50
328
730
(64,21%)
2.048

1.024
1.024
3.072
1.024
1.024
3.072
1.536
2.048
1.024
1.024
1.536
2.048
1.024
1.024
43
43
148
(13,01%)
786
1.137,0
Ket qua 3 dgt san
xuSt
huyet thanh Sdi, 10 thd cho
san
xuat dgt 1, 10 thd cho
san
xuit dgt 2
va
9 thd cho
san

xuit dgt 3. Hifu gia khang huyft thanh tir
Idn
hon
1000 don vi sg sir dung cho
thii
nghifm trung hda me gat don phat hien tac nhan ngdai
lai.
Trong dgt
1 san
xuit huylt thanh, 3 thd cd khang huyet thanh
hifu
gia thip
nSn
khdng
sii dyng
dugc, 7/10 cd khang huyet thanh cao hon dugc su dung. So vdi khang
huylt thanh
Nhjt
ban cung cip (hifu gia >4000 don vi) thi trong dgt
1
chi
4/10
thd dat
yeu ciu(l,2,4,7) hifu gia huylt thanh tir 4096-8.192; nhu vay chi 134/327ml huylt
14
thanh
san
xuat cd hifu gia khang huylt thanh tuong
duong
vdi

mlu
chuin cua Nhat
Ban trong
san
xuit dgt
1;
128/327 ml huylt thanh
san
xuit cd hifu gia thip hon miu
chuan Nhat ban nhung dat tieu chuin eg sd; 105/327ml huylt thanh san xuit ra khdng
dat tifu chuin co sd
Trong dgt 2,
10/10
thd
sii
dung dugc huylt thanh san xuit ra, tuy nhien chi cd
2/10 thd
djt
hifu gia tuong ducmg vdi huylt thanh Sdi do Nhat Ban cung cip (thd
30,33) vdi hifu gia 6.144, 8.192. Nhu vay chi 80/397ml huylt thanh san xuit ra tuong
duong vdi huylt thanh Sdi chuin;
Trong dgt 3, huylt thanh cua 9/9 thd dugc sir dying cho kilm dinh
vie
xin Sdi,
tuy nhifn chi 1/9 huylt thanh thd dat tuong duong huylt thanh chuin Nhat ban(42) vdi
hifu gia huylt thanh
6.144,
Nhu vay dgt 3 chi cd 45/373ml huylt thanh dat tuong
duong vdi huyet thanh
nhjp

ngoai.
Nhu vay khang huylt thanh Sdi dugc san xuit,
hifu
gia dat tieu chuin Nhat ban
chilm
22,78%; Huylt thanh dat hieu gia tieu chuin co sd
64,21%
va
13,01%
lugng
huyft thanh thap khdng dat tieu chuin co sd qui dinh.
Khang
huygt
thanh Sdi dugc dung cho cac thd nghifm
kim
dinh vacxin thanh
pham (Chuan
dd
hifu gia
va
dn dinh nhift
;Thii
nghifm nhan dang);kifm dinh vacxin
ban thanh
pham(Chuin
dg hifu gia
va
dn dinh
nhift;Thii
nghiem tac

nhSn
ngoai lai tren
3
loji
te
bao:
Vero, PL, CEC: The tich mlu:
50
ml cho mdi loai te bio; Trung hda theo
ty
If
1:1; Ndng
dd
khi
hrung
hda 480 don vi khang huylt thanh);
Thii
nghiem tac nhan
ngoai lai trfn tning ga cd phdi: tiem Allantoic va Yolk sac
(Thi
tich miu:
11
ml cho
mdi dudng
tiSm;
Trung hda theo ty
If
1:1; Ndng dg khi trung hda 480 don vi khang
huylt thanh)
Do sd lugng huyft thanh thd sir dung rat

Idn
khi
san
xuit 7,5 trifu lilu vie xin.
Vif c
cSn
thiet phai
thiSt lap
qui trinh san xuat khang huyet thanh Sdi cd hieu gia cao
va
dong deu tren thd sir dung. Hifn nay
POLYVAC
dang
thilt
lap qui trinh san xuit
khang nguyen vk chgn
Igc
thd de dat dugc ket qua nhu
y
mudn.
3.2.
Khing
huyet thanh
B^i
liet
Huyet
thanh
B?i
lift
typ

Sd
thd
Thi
tich
(ml)
Hieu gia Sir dung
trung
hda
me
gat don
(ml)
Hf sd
pha loang
POLYVAC
Nhat
ban
Chuan dp
hifu gia
vac xin tam
lien
(ml)
/
Tdng
1
2
Tdng
2
3
T6ng3
T6ng

c$ng
1
2
3
4
5
7
9
10
8
tho
2
3
4
6
7
8
9
7
thd
11
12
14
16
17
21
23
7
thd
29

thd
33
40
38
25
35
28
33
44
276
45
41
25
41
28
21
26
227
53
35
35
40
23.5
29
25
240,5
743,5
262144
1048576
16384

262144
65536
131072
65536
131072
512
2048
8192
2048
2048
512
512
16384
16384
4096
2048
2048
64
512
x
X
X
X
Cheo
typ 2
tai
ty
If
pha
loang

128
X
X
X
170
X
25
X
X
88 ml
283
512
2.048
512
256
256
16
32
32
128
128
128
128
128
32
64
X
Cheo
typ 2
X

71
X
Cheo
typ 3
X
X
Cheo
typ 3
X
140
X
X
X
X
Cheo
typ 2
127,5ml
338,5
Ket qua san xuit huylt thanh Bai lift typ
1
tren 8 thd vdi tdng huylt thanh thu
duoc 276 ml vdi hifu gia rit cao.5/8 thd cd hieu gia khang
thi
cao nhit,
thi
tich huylt
thanh 170ml su dung cho thu nghifm trung hda me gat don, hf sd pha loang
tii
256-
2048 lan, vdi ty

If
pha loang nay, lugng huylt thanh se sir dung dugc it nhit trong
vdng 20 nam cho
thii nghifm
tac nhan ngoai lai. 2/8 thd vdi hieu gia thip hon
sii
dung
cho kiem tra hifu gia
vie
xin tam lien vdi ndng
do
pha loang
1
don
vi/ml.
1/8 thd huylt
thanh bi loai vi cheo vdi typ
3
tai ndng do pha loang 128 lin.
Hieu gia khang the typ2 thip hon rit nhilu so vdi typ
1.
Chi cd 1/7 thd, hifu gia
huyet thanh cd
thi
tnmg hda hoan toan me gat don vdi ty
If
pha loang thip (16 lin).
4/7 thd cd
hifu
gia huyet thanh thap

hem
nhung rit dac
hifu
du tieu chuin su dung cho
thii nghifm
xac dinh hieu gia
vie
xin Bai het tam lien vdi ndng do
1
don vi /ml.
Hifu gia khang
thi
typ
3
san xuit tren 7 thd, mac
dii
hifu gia khang
thi typ3
cao
hon hifu gia khang
thi
typ 2 nhung
vin
thip hon
hifu
gia khang
thi
typ
1
rit nhilu.

Chi 2/7 thd cd huyft thanh dat dugc tieu chuan cho thir nghiem xac dinh tac nhan ngoai
lai trong me gat don vac xin vdi ty
If
pha loang 32 lin. 4/7 thd, huylt thanh chi sir
d\mg
cho xac dinh hifu gia trong
vie
xin thanh phim. 1/7 thd khdng
sii dimg
dugc
huyft thanh do cheo vdi typ 2.
Nhu
v?y
qua 22 thd sir dung cho
san
xuit khang
thi
Bai lift typ 1, 2, 3 ta thiy
ring so lugng
thi
tich khang thf thu dugc 743 ml trong dd hieu gi dat va cao hon
khang
thi
chuan
ciia
Nhat
Ban la
283nil,
khang the nay
sOf

dung cho
thii
nghifm tac
nhan ngoai lai bing phuong phap trung hda hoan toan me gat don. 122
ml
khang
thi
bi
lo?i
bd vi khdng dac hifu
typ;
khang the cdn lai su dung cho thir
nghifm
tim hieu gia
trong
vie
xin thanh phim vdi
sir
dung
1
don vi /ml.
Ciing qua ket qua tren khang the Bai lift typ
1
rit cao, cao hon rat nhilu so vdi
khang the Bai lift typ 2 va 3 can nghien
ciiu
san xuit khang the typ 2, 3 sao cho hifu
gia cao nhu
typl
de

ling
dung cho thir nghifm xac dinh tac nhan ngoai lai trong me gat
don.
3.3.
Khang huyet thanh Rota
The tich
va ling
dung
Sdtube
Ung dung
™^'
ChOngGN ^ ^Jf
Hifugia
^f
Tylepha
mVtube) loang
'^^'^
xac
dinh
17
hieu gia
1
3
4
5
6A
7
8A
9
10

6
18
8
22
13
20
24
25
TSng
cOng
G1P8
G1P8
G1P8
G1P8
G1P8
G1P8
G1P8
G1P8
G1P8
Wa
Wa
DSl
DSl
RRV
RRV
RRVmix
RRVmix
42
44
35

48
35
42
50
47
55
398
45,0
39,0
84
51,0
39,0
90
60,0
48,0
108
36,0
25,5
61,5
741,5
27
27
20
27
20
25
30
27
32
235

30
26
56
34
26
60
40
32
72
24
17
41
974
102400
102400
25.600
102400
6.400
102.400
25.600
25.600
6.400
1.638.400
1.638.400
409.600
409.600
1.638.400
1.638.400
1.638.400
409.600

1:
1:
1;
1:
3.000
3.000
3.000
1.000
1.10.000
1.10.000
1:2.500
1:10.000
1:5.000
1:10.000
1:2.500
1:2.500
1:5.000
1.30.000
1.30.000
1.10.000
1.10.000
1.30.000
1.30.000
1.30.000
1.10.000
Tong sd khang huyet thanh sdn xuat dugc 741,5ml trong dd chung G1P8
(398ml), WA (84ml); DSl
(90ml);
RRV (108ml)
va chiing

RRV mix
(61,5ml)
Hifu
gia huylt thanh Rota
sdn
xuit
tii
5 chiing deu cho hieu gia rat cao thap nhat
chiing G1P8 (6.400)
Hifu
gia huylt thanh Rota
san
xuat tir chiing hoang dai cao hon chung giam ddc
lyre
rit nhilu, hifu gia chiing G1P8 giam dgc
lire
chi bang
'A
den 1/16 chung
hoang dai
Huylt thanh san xuit tit chung RRV dugc ling dung tdt trong thir nghifm tun
hifu gia
vie
xin Rota vi
hhih
anh
kit
qua r6 net va dep; Huylt thanh
tii
nhiing

chiing khac sir
d\mg
cho thtr nghifm dimg
ky
thuat ELISA.
KET
LUAN
1.
Da ling dung
va
hoan thien qui trinh san xuat huyet thanh Sdi, Bai
lift.
Rota
sii
dung tdt cho kiem dinh vac xin
• Qui trinh san xuit huylt thanh Sdi
San xuat khang nguyen tren te bao vero
Tinh sach khang nguyen va pha khang nguyen vdi ndng do
lO'
CCID/0,5ml
Gay dap
irng
mien dich tren tho
sir
dung ta
diroc
Thuc hifn 5 lan gay
dap
img miln dich theo lich ngay: 0 : 7 : 14: 28 : 42. Lilu lugng
tiem mdi lan nhu sau:

TiSm Ik
1
ng4y
0; TiSm 2,5ml
h5n
djch virut + 2,5
ml ti
chit FCA;
Tifim
trong da
nhi^u
vj tri,
0,1
mI/1
vi tri;
TiSm
lan 2 ngiy 7: Tiem 5 ml virut Soi khdng c6
ti
chit; TiSm
vio
dudi da
diii
2
bSn,
m5i bSn
2,5 ml;
Tiem lan 3
ngiy
14'
TiSm 5 ml virut Soi khdng c6 ta chit; Tiem

v4o duoi
da
diii
2
bSn,
mfli
bSn 2,5 ml;
TiSm lin 4
ngiy
28:
TiSm 10 ml virut Soi khdng c6 ta chit; TiSm
vio du6i
da
dCii
2 bSn, mdi bSn 5 ml;
TiSm lin 5
ngily
42; TiSm
10
ml virut Sdi khdng cd tii chit; TiSm via
tinh
mach tai 2 bSn, mdi bSn 5 ml
Lay mau, tach huyet thanh
Lay mau
vao
ngay
thii
49 sau tiem lan
1,
tach huyet thanh

Chuan
d$
hifu gia, xac dinh dg dac hifu
Chia nhd va bao quan
Qui trinh
san
xuat huyet thanh Bai
lift
San xuat khang nguyen tren te bao vera
Tinh sach khang nguyen va pha khang nguyen vdi nong do 10 CCID/0,lml
Gay dap
irng
mien djch tren thd sir dung ta dugc
Thd dugc each ly
tii
7-10 ngay, gay
dap ung
miln dich 6 lan tiem
each
nhau 2 tuan vao
ngay:
0:14:28:42:56:70;
84.
Lich va Ulu
tiem cu
thi:
TiSm lin
i ngiy
0: TiSm 0,2ml virut Bai liet + 0,2ml ta
dupe

FCA; Tiem trong da
nhiSu
vi tri, 0,1 mi/1 vi tri;
TiSm lan 2
ngiy
14: TiSm
Iml
virut Bai liet + 1ml ta
tlupc
FCA; Tiem
v^
CO
diii 2 ben, mdi ben 0,5 ml;
TiSm lin 3
ngiiy
28:
TiSm 1ml virut Bai list + Iml ta dugc FCA; TiSm
vio
co
diii
2 bSn, mdi bSn 0,5 ml;
TiSm lin 4
ngSy
42:
TiSm 1ml virut Bai
liSt
+ 1ml
ti dirgrc
FCA; TiSm
vio

co
diii
2 bSn, mdi ben 0,5 ml;
TiSm lin 5
ng4y
56: TiSm 1ml virut Bai
liSt
+ 1ml ti duoc FCA; TiSm vao co
diii
2 ben, mdi ben 0,5 ml;
TiSra lin
6 tlgiy
70: TiSm 1ml virut Bai
liSt
+
Iml
ta duoc FCA; TiSm
v&o
co
diii
2 bSn, mdi ben 0,5 ml;
Lay mau, tach huyet thanh
Lay mau
toan bd cudi
cimg vao
ngay 84 sau lin tiem 1
Chuan dg hifu gia, xac dinh dg dac hifu
Chia nhd va bao quan
• Qui trinh san xuat huyft thanh Rota
San xuat khang nguyen

tr£n
te bao vero
Tinh sach khdng nguyen
va
pha
khing
nguyen vdi nong dg 10
CCID/lml
GSy
dap irng mien djch tren tho sir dung ta
dirge
Thd dugc each ly
tii
7-10 ngay, gay dap iing mien dich 4 lan tiem each nhau 2 tuan vao
ngiy:
0:14:28:42 . Lich va lilu tiem cu the:
Tiflra
lin I
ngiy
0: Tiem 0,2ml virut Rota + 0,2m! ta
dugrc
FCA; Tiem trong da
nhik
vi tri, 0,1
mJ/1
vj tri;
Ti6m lin 2
ngiy
14: TiSm 1ml
vinit

Rota + 1ml ti
dirijc
FCA; Tiem
v4o
ca
diii
2 ben, m5i
b8n
0,5 ml;
TiSm lin 3
ngiy
28: TiSm 1ml virut Rota + 1ml
ti dtrpc
FCA; TiSm vio co
diii
2
bdn,
m6i
ben 0,5 ml;
Tifim
lin 4 ngiy 42: TiSm 1ml virut Rota + 1ml ta
du(7c
FCA; Tiem
vio
ca
diii
2 ben,
m6i
ben 0,5 ml;
Lay man, tach huyet thanh

Liy
mau
toan bd cudi cimg
vao
ngay
56
sau lan tiem 1
Chuan do hieu gia, xac dinh do dac hieu
Chia nhd va bao quan
2.
Da san xuat dugc huyet thanh Sdi, Bai
lift.
Rota dat tifu chuan co sd
Huyet thanh S&i
Trong
1.097nil
huylt thanh san xuit dugc (327 ml dgt 1, 397ml dgt 2
va 373
ml
&qt
3) cd 259ml huylt thanh cd hifu gia tuong duong vdi miu chuin Nhat ban (Hifu gia
>4000 don
vi);
105 ml huyet thanh hifu gia qua thap khdng
sii
dung dugc; lugng huyet
thanh cdn lai mac du khdng cao
bing
huyet thanh chuan nhung van
sii

dung dugc trong
kiem dinh vie xin Sdi.
Huyet thanh Bai lift
Sd lugng
thi
tich khdng huylt thanh Bai lift
typ
1, 2, 3 thu dugc
743
ml (276 ml
huylt thanh typ 1, 227ml huylt thanh Bai lift typ 2 va 240,5ml huylt thanh Bai lift
typs 3) trong dd hifu gia dat
va
cao hon khang huylt thanh chuin ciia Nhat Ban la
283ml chu
ylu
la huylt thanh typ l(170ml), khang huyet thanh
niy sii
dvmg cho thir
nghifm tac nhan ngoai lai bing phuong phap trung hda hoan toan me gat don. 122 ml
khang huylt thanh bi loai bd vi khdng dac
hifu
typ;
khang huylt thanh cdn lai
sii dimg
cho
thu:
nghifm
tim hieu gid trong
vie

xin thanh phim vdi
sii
dung
1
don vi /ml.
Huyit
thanh Rota
T6ng sd khang huylt thanh
san
xuit dugc 437,5ml trong dd chung G1P8, WA
(84mV
chimg)
; DSl ( 90ml); RRV (108ml) va chiing RRV mix (61,5ml); Hifu gia
huylt thanh Rota san xuat tir chung hoang dai cao hon chiing giam dgc
lire
rat nhieu,
hifu gia chiing
G1P8
giam dgc
luc
chi bang
'A
dfn
1/16
chiing hoang dai; Huyet thanh
sdn
xuit tit chiing RRV dugc iing dung tdt trong thu
nghifm
tim
hifu

gia
vac
xin Rota
vi hinh
dnh kit
qua ro net va dep; Huyet thanh
tii
nhiing chiing khac su dung cho thir
nghifm dimg ky thu$t ELISA.
3.
Khang huylt thanh da dugc thi nghifm img dung kiem tra trong thii nghifm
tac nhan
ngo?i
lai,
thii nghifm
nhan dang, thir
nghifm xac
dinh
hifu
gia vac xin, trong
ky
thu|t
ELISA so vdi
mlu
khang huylt thanh nhap ngoai cho ket qua tot, thay
thi
dugc khang huylt thanh
nh|p
ngoai
KIENNGHI

1.
Dugc sir
dvmg
khang huyet thanh
nay
trong kilm dinh vic xin thay
thi
khang
huyet thanh nhap ngoai;
2.
Nghien
ciiu
tiep tuc tinh dn dinh cua san pham
tao
ra;
3.
Nghien
ciiu
qui trinh
san
xuat khang huyet thanh Sdi,
Bsii
lift
typ
2,3 sao cho
cd
san
pham dat
hifu
gia cao hon.

Bai
bao
va dao
tao
Bai
bao
da dang:
01
bai
Nghien
ciiu
san xuat huylt thanh Sdi
sii
dung cho
Idem
dinh vac xin.Tap Chi Y
hgc
dvr
phong,
2008, Tap
XVIH
sd 6(98), tr.74-78
Dio
tgio:
01
thacsy
Ten de
tai:
Nghien
ciiu

san xuat khang huyet thanh
Bai
lift trfn thd
va
danh gia
tinh an
toan
cua vie xin
Bai
lift trong phong thi nghifm.
Chuyf n
nhtoh:
Thii
y
Ma sd 60.62.50
Da
bao vf vdi
dilm
xuit sic
nim
2008
MUC
LUC
DAT VAN
DE 1
CHUONG1.
T6NG
QUAN 2
1.1. Khang nguyen 2
1.1.1. Tinh sinh mien

tlich va
tinh khang nguyen 2
1.1.2.
Cac yeu to anh
hudug din
tinh sinh miln dich 2
1.1.2.1. Nhiing tinh chat cua ban than khang nguyen anh hudng den tinh sinh mien
dich 2
1.1.2.2.
Nhung tinh chat cua he thong sinh hoc anh hudng den tinh sinh mien dich 4
1.1.2.3.
Quylt
dinh khang nguyen 7
1.1.2.4.
Khang nguyen vi sinh vat 7
1.2.
BChang
huylt thanh 8
1.2.1.Cau
tnic
CO
ban cua khang huylt thanh 9
1.2.2.
Cac quyet dinh khang nguyen tren phan tir globulin miln dich
11
1.2.3.
Cac l(5p
khang huylt thanh 13
1.2.4.
Ciu true

khang huyet thanh 18
1.2.5.
Ai
luc
cua khang huylt thanh vdi khang
nguySn
19
1.2.7.
Vai tro cua
khing
huylt thanh 20
1.2.8.
Khang huylt thanh
dcm
dong
va
da dong 21
1.3. Co chl hinh
thinh
khang huylt thanh 22
1.3.1.
cac khainifm
22
1.3.2.
Lieu lugng
va
dudng vao cua khang nguyen 23
1.3.3.
Cdc
te

bao
tham gia
vao
dap img mien djch
t^o
khang huyet thanh 23
1.3.4.
Ta
chat, nguyen ly tac
dyng
cua ta chit 24
1.3.5.
Co chl
hinh
thanh
khing
huylt thanh 24
1.4. Khdng nguyen virut 24
1.4.1.VirutSdi
24
1.4.1.1.
Ciu
talc 24
1.4.1.2.
Qua trinh
iMn
len cua vinit Sdi 26
1.4.1.3.
Dap ung mien dich 27
1.4.2.

Virat Rota 28
1.4.2.1. Ve
hhih
thai hpc 28
1.4.2.2.
Vl
thanh phin hoa hpc cua
vmit
Rota 29
1.4.2.3. vl tinh chit hda ly 29
1.4.2.4.
Ve cau
t?o ciia
khang nguyen 29
1.4.2.5.VI
tinh smh miln dich 30
1.4.2.6.
Vl
su nhan len cua virat Rota
frong
te bao cam thu 30
1.4.2.7.
vl virat Rota giam dpc luc dong Sabin 32
1.4.3.
Vuut
Rota 33
1.4.3.1. Phan loai vuut Rota 33
1.4.3.2.
D§c diSm
cua virat Rota 33

1.4.3.3. Hinh
thdi
va ciu true 34
1.4.3.4.
cac
thanh phin hoa hpc cua vuut Rcta
va chiic
nang 34
1.4.3.5.
Phan nhdmva typ
huylt thanh 35
1.4.3.6.
Tmh chat hoa ly 36
1.4.3.7.
Nuoi
ciy
._.
37
1.4.3.8.
Sv
nhan len cua virat Rota trong tl bao 37
1.5.
Thd 39
l.S.l.DSctinhsmhhpccuathd
39
1.5.2.
Cac loai thd dugc nuoi va sir
dung
tai
nude

ta 40
1.5.2.1. Dac tinh chung 40
1.5.2.2.
Phdnlosd
40
1.5.2.3.
Chon gidng
thd 41
1.5.2.4.
Chuong
nuoi 42
1.5.2.5.
Thtrc an
43
1.5.2.6.
Cham
sdc 43
CHUONG
II
45
NGUYEN VAT
LlEU
VA PHUONG PHAP NGHIEN CUU 45
2.
I.Nguyen
vat lieu va phuong pliap san
xuit khang nguyen 45
2.1.1.
Nguyto
vat lieu va phuong phap san xuit khang nguyen Sdi 45

2.1.2.
Nguyen vat
Ufu
va phuong phap san xuit khang nguyen Bai Het 46
2.1.3.
Nguy6n vdt
lieu va phuong phap san xuit khang nguyen Rota 47
2.2.
Nguyen vat
lifu
va phuong phap san xuit khang huylt thanh 48
2.2.1.
Nguyen vat lifu
va
phuong phap san xuit khang huylt thanh Sdi 48
2.2.2. Nguyen vit lifu vk phuong phap san xuit khang huylt thanh Bai lift 51
2.2.3.
Nguyen
vkt
lifu
va
phuong phap
san
xuit khang huylt thanh Rota 52
2.3.
xac
dinh hifu gia khang huylt thanh 52
2.3.1.
Xac dinh
hifu

gia khang huyet thanh Sdi 52
2.3.1.1.
Nguyen v$t
lifu 52
2.3.1.2.
Phuong phap
xac
dinh hieu gia khang huylt thanh Sdi 54
2.3.2.
xac
dinh hifu gia khang
huySt
thanh Bai Hft 59
2.3.2.1.
Nguyen vat lifu 59
2.3.2.2.
cac
budc tiln
hdnh
59
2.3.
3.
xac
dinh hifu gid khdng huylt thanh Rota 62
2.4. Kilm tra v6 khuin khdng
huySt
thanh 62
2.4.1.Vjt
lifu vd dung
cy

62
2.4.2. Cdc budc tiln hdnh 63
2.4.2.1.
Chuan bi
Ing thii
63
2.4.2.2. Tiln hdnh thir 63
2.4.2.3.
Dpc
kit
qud 64
2.4.2.4. Ddnh gid
kit
qua 64
2.5.
iTng
dung khang huyet thanh trong kiem dinh vac xin 65
2.5.1.
xac
dinh vuut ngoai lai (khdng huyet thanh Sdi, khdng huyet thanh Bai
Hft)
65
2.5.2. Xac dinh tinh ddc hifu, hieu gid cua virat Rota
bSng
phuong phap miln dich
huynh quang (dp dung cho khdng huyet thanh Rota) 65
2.5.2.1.
Nguyen
vdt Hfu
nghien

ciiu
65
2.5.2.2. Phuong phap tien hanh 66
2.5.2.3.
Dgc
kit
qud 66
2.5.3.
Sii
dung huylt thanh trong ky thuat ELISA (khdng huylt thanh Rota) 67
CHUONG
111.
KfiT
QUA VA BAN LUAN 68
3.1.
Qui trinh sdn xuat khdng huylt thanh tren thd 68
3.1.1.
Qui trinh sdn xuit khdng huylt thanh Sdi 68
3.1.2. Qui trinh sdn xuat khdng huyet thanh Bai Het 69
3.1.3.
Qui trinh sdn xuat khdng huylt thanh Rota 72
3.2.
Kit
qud sdn xuit khdng huylt thanh 73
3.2.1.
Kit
qua san xuat khang huyet thanh Soi 74
3.2.2.
K6t
qua hieu gia khang huyet thanh gay dap ling mien dich lan

1
74
3.2.3.
Ket qua hieu gia khang huyet thanh lan 2 gay dap ung mien dich 76
3.2.4. Ket
qua
hieu gia khang huyet thanh lan
3
gay dap ung mien dich 77
3.2.5.
Tinh
hinh
sir dung khang huyet thanh Soi 78
3.3.
Kit qua san xult
khang huylt thanh Bai het 80
3.3.1.
S^
xuit khang huylt thanh Bai liet typl 81
3.3.2. Ket qua san
xuSt Idiang huySt
thanh Bai het typ 2 85
3.3.3.
Kit
qua hieu gia khang huylt thanh Bai Het typ
3
90
3.3.4. Ung dung trung hoa hoan toan me gat don 94
3.4.
Kit

qua san xuit khang huylt thanh Rota 96
3.4.1.
Ket qua khang huyet thanh san xuat lan
1
(khong chon tho truac khi tiem) 96
3.4.2.
Kit qui
khang huylt thanh san xuit
l§n
2 98
KETNGHI
107
TAI
LI$U
THAM KHAO 108