Mc lc

Trang
M U...............................................................................................................................................................1
Chng 1 - TNG Q U A N ................................................................................................................................ 3
1.1. Virut s i....................................................................................................................................................3
1.1.1. Phn loi............................................................................................................................................3
1.1.2. Hnh thi h c ................................................................................................................................... 4
1.1.3. Cu trc khng nguyn.................................................................................................................. 4
1.2. Bnh s i....................................................................................................................................................5
1.2.1. Chu k pht trin ca virut si trong c th bnh nhn..............................................................5
1.2.2. p ng min dch t nhin vi virut s i.................................................................................... 7
1.2.3. Bin chng ca bnh s i................................................................................................................ 8
1.2.4. Tnh hnh mc bnh si hin n a y ..................................................................................................9
1.2.4.1. Tnh hnh mc bnh si trn Th gii................................................................................... 9
1.2.4.2. Tnh hnh mc bnh si Vit N am ....................................................................................11
1.3 Phng v iu tr bnh s i................................................................................................................... 14
1.3.1. Vacxin phng bnh s i.................................................................................................................14
1.3.1.1. Vacxin bt hot.......................................................................................................................15
1.3.1.2. Vacxin sng gim c lc.....................................................................................................15
1.3.1.3. Vacxin si kt hp vi vacxin quai b v rubella (M M R )............................................... 16
1.3.2. iu tr bnh s i............................................................................................................................ 17
1.4. Tnh hnh tim chng vacxin si phng bnh ti Vit N am ............................................................ 17
1.5. Sn xut vacxin si ti Vit N am ........................................................................................................ 18
1.6. Kim nh vacxin si sn xut ti Vit N am .....................................................................................19
1.7. Th nghim lm sng........................................................................................................................... 19
Chng 2 - VT LIU V PHNG PHP NGHIN c u ................................................................. 21
2.1. Phng php chun hiu gi bng to m hoi t ..................................................................... 21
2.1.1. Nguyn vt li u ..............................................................................................................................21
2.1.1.1. Mu chun.............................................................................................................................. 21
2.1.1.2. Ho cht, mi tr ng............................................................................................................ 21
2.1.1.3. Dng c ...................................................................................................................................21

2.1.1.4 T bo s d n g .......................................................................................................................22
2.1.1.5. Mi trng s d n g ..............................................................................................................22
2.1.2. Quy trnh thc hin.......................................................................................................................24
2.1.2.1. Chun b mu v mu ch u n ...............................................................................................24
2.1.2.2. Pha long, gy nhim, n ui................................................................................................. 24
2.1.2.3. Phng php tnh kt qu.....................................................................................................26
2.2. Phng php kho st hiu gi vacxin si dng cho th nghim lm sng..................................28
2.2.1. Phng php ly m u .................................................................................................................. 28
2.2.2. uy trnh chun hiu g i ........................................................................................................ 29
2.2.3. Phng php kho st.................................................................................................................. 29
Chng 3 - KT QU V BN LU N.....................................................................................................30
3.1 Ket qu hiu gi vacxin si (M-0107)................................................................................................ 30
3.2 Kt qu hiu gi vacxin si (M-0207)................................................................................................ 32
3.3 Ket qu hiu gi vacxin si (M-0307)................................................................................................ 34
3.4 Ket qu hiu gi vacxin si i chng (R )........................................................................................ 36
KT L U N ..................................................................................................................................................... 39
TI LIU THAM KHO...............................................................................................................................40

M U
Bnh si l mt bnh nhim trng cp tnh do virut si gy nn, ly nhim theo
ng h hp v d ly thnh dch, c bit l tr em. Bnh si tuy c nhn bit
t hn 2000 nm trc y, nhng n nm 860 Zanes mi phn bit bnh si khc
vi bnh u ma [7]. Vi thp k gn y, Y hc hin i c nhiu thnh tu trong
vic phng tr bnh dch. Mc d vy, cho n nay bnh si vn cn l mt vn
dch t nghim trng. c tnh hng nm trn Th gii c khong 50 triu trng hp
nhim bnh v trn mt triu trng hp t vong lin quan n si [8,9].
Nm 1954, nhng thnh cng ca Enders v Peebles trong vic phn lp virut
si nh du mt bc tin mi trong nghin cu bnh si. Vic nghin cu dch t
hc bnh si c thc hin trn nhiu nc v kt qu cho thy vacxin si c vai
tr then cht trong vic phng bnh si. Cc chin dch tim chng trn din rng v
phc tim vacxin lm gim ng k t l mc bnh si.
Tuy nhin trong nhng nm gn y dch si li bng n nhiu nc nh M,
Anh, Canada, ti cc nc ang pht trin, c bit l cc vng nhit i chu Phi v
chu . Vic phng chng bnh si cn gp nhiu kh khn, dch bnh vn cn honh
hnh ng k. Vit Nam, bnh si vn cn l bnh nguy him i vi tr em nm
2000 c 16.200 tr em mc bnh si [2]. V th t nhng nm 1982 vacxin si l
mt trong

vacxin c a vo chng trnh tim chng m rng gy min dch

ch ng cho tr em. Mc tiu cui cng ca chng trnh ny l thanh ton bnh si
trn ton Th gii. Mun thc hin c hiu qu chng trnh tim chng th vic m
bo cht lng vacxin, trong c hiu lc ca vacxin l ht sc quan trng.
Hiu lc ca vacxin si c th hin bng hiu gi virat v c xc nh bng
kh nng gy hy hoi t bo cm th Vero ca virut vacxin. Hiu gi l ch tiu quan
trng quyt nh tnh sinh min dch v hiu qu bo v ca vacxin. V l vacxin sng
gim c lc, rt nhy cm vi nh sng v nhit nn hiu gi ca vacxin si d b
1

nh hng trong qu trnh bo qun v vn chuyn n ngi s dng nht l trong

iu kin kh hu nhit i ca nc ta, do ngoi vic kim tra hiu gi ca vacxin
khi mi sn xut ra cn phi gim st kim tra cht lng, hiu gi ca vacxin trn
thc a.
e gp phn vo vic nh gi cht lng vacxin dng trong chng trnh tim
chng m rng ti Vit Nam, chng ti thc hin ti nghin cu: Kho st hiu
gi ca vacxin si c sn xut ti Polyvac dng cho th nghim lm sng. Nhm
gii quyt 3 mc tiu:
1. Hiu gi vacxin Si sn xut ti Polyvac t tiu chun ca WHO
2. Hiu gi vacxin ti bn tim th nghim lm sng cng t tiu chun ca
WHO
3. Bin ng hiu gi vacxin trong qu trnh vn chuyn vacxin ti a im tim

Chng 1 - TNG QUAN

1.1. Virut si
1.1.1. Phn loi
Virut si l mt trong 3 thnh vin trong chi Morbillivirut thuc h
Paramyxoviridae. H ny gm 3 nhm
- Cc Paramyxovidae:
+ Para - influenza 1, 2, 3, 4
+ Virut quai b
+ Virut gy bnh Newcastle
+ Virut Sendai
- Cc virut Pneumo: Virut hp bo ng h hp
- Cc virut Morbilli:
+ Virut si
+ Virat gy bnh Carre ch
+ Virut dch hch b
Ba virut thuc chi Morbilli c mi lin quan cht ch vi nhau nhng
im sau:
- Ging nhau v hnh thi, kch thc
- Chng c c im chung ni bt l u gy hiu lc hy hoi trn nui
cy t bo to cc hp bo a nhn, cc tiu th trong bo tng v trong nhn
- Ba virut ny l cc tp n. u gy bnh ging nhau trn vt ch mn
cm [2 0 ]

1.1.2. Hnh thi hc

Nhng kt qu nghin cu dng siu ly tm v knh hin vi cho thy cc
virion si l nhng ht c v bc, hnh cu, vi li nucleocapsid hoc l xon
cun hoc l hnh si ch. ng knh c nhiu kch c, thay i t 120-150 nm,
tnh trung bnh l 150 nm bao gm

protein. Bn trong cav capsid l chui

ARN l b gen di truyn c cu to hnh xon cvi 3 protein, v ngoi cng

ca virut si l chat glycoprotein vi cht ngng kt hng cu (H) hnh nn l
mt khng nguyn ly nhim quan trng ca virut si v protein F [1].
1.1.3. Cu trc khng nguyn
Norrby v Gollman miu t 4 cu trc khng nguyn khc nhau ca
virat si:
- Khng nguyn kt hp b th
- Khng nguyn ngng kt hng cu
- Khng nguyn tan mu
- Khng nguyn trung ha
Trong , hai khng nguyn chnh nhn bit virut si l khng nguyn
ngng kt hng cu v khng nguyn tan mu [ 11 ].
Cc nghin cu cho thy khng nguyn gy ngng kt hng cu l khng
nguyn gy p ng min dch bo v v khi tim khng nguyn ngng kt hng
cu tinh khit cho ngi v ng vt thy xut hin khng th si vi hiu gi
cao, ging nh gy min dch bng vacxin si bt hot bi formalin [4].
S tc ng gia khng nguyn ngng kt hng cu ca virut si vi hng
cu kh ch yu l do phn ng enzyme v tc phn ng ngng kt hng cu
tng khi nhit tng n 37c [3]. Rt c th ban u enzyme ca virut ph

hy mt s cu trc trn b mt hng cu, sau cc th th c bc l, to

iu kin gy ngng kt hng cu.
Khng nguyn ngng kt hng cu bn vng vi nhit 60c trong hn
1 gi, nh c im ca vi rt si khng c hot tnh neuraminidaza [5].
Cc nghin cu cho thy cc th th do hot tnh ngng kt hng cu
ca vi rt si khng b ph hy bi cc enzyme ph hy th th. Cc khng
nguyn ngng kt hng cu cng khng nhy cm vi cc enzyme ARN-aza
v ADN-aza [7].
Khng nguyn tan mu l mt glycoprotein c trn b mt v ngoi virut
vi chc nng dung hp t bo to cc hp bo. Hot tnh tan mu ca virut
si c lin quan cht ch vi kh nng gy hy hoi t bo sm, n gip cho
virut si d dng xm nhp vo bn trong t bo v tng ng vi kh nng gy
nhim ca virut.
Khng nguyn tan mu c pH thch hp l 8,0 [14].
Khng nguyn trung ha nm v ngoi virut v tham gia vo phn

ng

trung ha [3,6].
1.2. Bnh si
1.2.1. Chu k pht trin ca virut si trong c th bnh nhn
Bnh si l mt bnh nhim trng ng h hp c kh nng ly nhim
cao. Bnh ny hay ly n ni 90% nhng ngi tip xc vi bnh nhn s b ly
si nu cha tim phng. Virut si c mi v c hng ca bnh nhn. H
thng c th ly bnh cho ngi khc 4 ngy trc khi vt xut hin. Khi
bnh nhn ho, ht x, hay ni chuyn, nhng git nc rt nh c cha virut s
bn ra khng kh v ngi khc c th ht vo v h s b ly bnh. Mt khi virut

si vo c th bnh nhn, chng thng nhn ln trong nhng t bo ng h

hp trn. Sau bnh s lan khp c th k c h h hp v da.
* Triu chng bnh: Khong 10 ti 12 ngy sau khi tip xc vi virut
si, nhng triu chng sau y c th xy ra:
- St, Ho khan, Chy nc mi, Mt , ....
- Nhng nt rt nh vi trung tm mu xanh trng xut hin bn trong
mim mc m. Nhng nt ny c tn l m Koplik.
- Ngi mc ra nhng m ln, phang, chm vo nhau
* Din bin ca bnh
Giai on bnh
Thi k bnh thng ko di t 10 n 12 ngy l thi gian t khi
virut gy bnh si xm nhp vo c th n khi xut hin nhng du hiu
u tin ca giai on tin triu. Trong giai on ny khng biu hin triu
chng g ca bnh.
Giai on tin triu chng
Giai on tin triu thng ko di 3 n 5 ngy c c trung bi st
mc nh n va, ho khan, chy mi nc, vim kt mc mt. Nhng triu
chng ny hu nh lun lun xy ra trc khi ni ban xut hin. Ni ban hay ht
Koplik l du hiu ch im ca bnh. Ht Koplik l nhng ht nh bng ht ct,
mu trng ng, xung quanh c vin . Ht thng xut hin v bin mt nhanh
trong vng 12 n 24 gi. Ket mc mt c th b vim v c du hiu s nh
sng.
Giai on pht ban

y l giai on in hnh nht ca bnh vi triu chng pht ban tun t

trn da. Ban thng xut hin u tin vng chn tc pha sau tai, sau xut
hin mt v lan dn xung pha di trong vng 24 n 48 gi. Ban si l
nhng ban dng dt-sn hi ni ln trn b mt da, s mn nh nhung v khng
au, khng hoc t nga, khng sinh m. Trong trng hp nh, ban thng
ng gn nhau nhng ring r. Trong trng hp nng, ban c xu hng hp vi
nhau lm thnh nhng ban ln hn, thm ch tng mng xut huyt (si en).
Trong th c bit nng, ban c th c du hiu xut huyt. Khi ban lan n chn
th st cng t ngt gim i nu khng c bin chng. Sau ban cng nht
dn v mt i ng theo tun t n xut hin, ngha l cng t trn xung
di. Sau khi ban mt i, trn da cn li nhng du mu sm lm m nh vn
da bo [ 10 ].
Si khng in hnh
Mt s trng hp bnh si biu hin khng ging nh miu t trn nh
trong trng hp bnh nhn b suy gim min dch, ban c th khng in hnh.
Cc trng hp ny thng thy bnh nhn AIDS, hi chng thn h, iu tr
thuc c ch min dch.
1.2,2. p ng min dch t nhin vi virut si
p ng min dch qua trung gian t bo l biu hin quan trng c giai
on bnh l v phc hi bnh, c ch p ng min dch c hiu bt u cng
vi nhng biu hin lm sng ca bnh, trc khi pht ban v ko di trong vng
vi tun sau khi hi phc bnh. Cc khng th do p ng min dch t nhin
ngay khi ban xut hin. Khi u l IgM tip sau l cc IgG v IgA trong huyt
thanh v dch tit. Cc khng th ny sinh ra nhm chng li cc khng nguyn
l trong c th chng t nh cao vo ngy th 7 n th 10 sau khi pht ban,

sau gim nhanh v bin mt. Cc khng th c sinh ra trc tip khng
li cc protein khc nhau ca virat c pht hin khng ging nhau bng cc
xt nghim huyt thanh hc. Khng th vi protein N c sn xut nhanh v
nhiu, c xc nh ngay giai on nhim trng tin pht bng k thut kt hp
b th. Chnh v s hnh thnh khng th khng N mc nhiu nh vy cho
php nhn nh kt qu l m tnh khi khng pht hin c chng trong huyt
thanh [7]. Sau khi khi bnh, c th chng ta s c khng th chng bnh nn ta
s khng mc bnh si li na. Neu bn sinh trc nm 1957, bn thng c
khng th chng bnh d cha h chng nga. l v bn sng qua nhiu
thi k m bnh si honh hnh rt nhiu v nhiu phn bn c nhim virut
d c th khng l ra. Do bn c c min dch.
1.2.3. Bin chng ca bnh si
Cc bin chng thng gp ca si l vim tai gia, vim phi, tiu chy,
vim no. Chnh cc bin chng ny lm ko di thi gian bnh, nh hng n
dinh dng ca tr. Hu qu l suy dinh dng.
1. Vim phi bin chng gy ra do chnh bn thn virut si (vim phi t bo
khng l). Vim phi do si bnh nhn AIDS thng gy t vong v him khi
c ban in hnh. Thng gp hn l bi nhim vi khun gy nn vim phi. Cc
vi khun thng gp l ph cu khun, lin cu khun nhm A, t cu khun v
Hemophilus Influenzae tp b.
2. Vim tai gia l bin chng lun lun phi ngh n tr mc si.
Nguyn nhn gy bnh cng tng t nh trong vim phi. Neu khng pht hin
kp thi, vim tai gia c th gy thng mng nh, nh hng n thnh lc. i
khi vim tai gia cp khng c iu tr ng c th a n vim tai gia mn
tnh vi bin chng nguy him l vim tai xng chm v p xe no.

3. Tiu chy cng l bin chng thng gp sau mc si, c bit nhng
tr suy dinh dng v thiu vitamin A. Tr d b mc l trc trng v tiu chy
ko di. i khi do c th suy kit, bnh nhi d c nguy c nhim trng huyt
tiu im t rut.
4. Vim lot gic mc: y l bin chng kinh in v ng s. Tr suy dinh
dng v thiu vitamin A l nhng i tng c nguy c cao nht. Bnh c th
din bin t lot gy m gic mc, hng ton b gic mc n lm m trong
nhn cu. Hu qu l gim th lc n m vnh vin ton b. Bin chng ny
hin nay cng gim r nh' iu kin dinh dng c ci thin v nh vo
chin dch b sung vitamin A cho cng ng.
5. Si lm suy gim chc nng min dch ca c th nn c th to iu kin
cho th lao tim n ti bng pht mnh m.
6. Vim c tim cng c th xy ra nhng him gp hn.
7. Vim no c tnh khong 1 - 2/1000 trng hp mc si. Khng c mi
tng quan gia mc nng ca bnh si vi kh nng xut hin vim no.
Cng khng c tng quan gia triu chng khi u ca vim no vi tin
lng ca n. C hai th vim no do si. Mt th l do phn ng min dch
thng qua s hnh thnh phc hp khng nguyn - khng th. Mt th khc l do
s hin din ca virut si tn ti trong t bo thn kinh gy nn vim no chm
c th xut hin 5 nm thm ch 15 nm sau khi mc si.
8.

Cc bin chng thn kinh khc l hi chng Guillain-Barre, lit na

ngi, huyt khi tnh mch no,... thng t gp [ 1 1 ].

1,2.4. Tinh hnh mc bnh si hin nay
1.2.4.1. Tnh hnh mc bnh si trn Th gii

Bnh si c mt khp cc nc trn Th gii. nhng nc tim phng

vacxin rng ri, t l mc bnh v t vong do si gim xung r rt, tuy nhin
trong nhng nm gn y, bnh si dn quay ch li v dch si bt u bng no
ngay c cc nc cng nghip pht trin nh M, H Lan, Anh, Trung Quc.
Theo thng k, k t nm 1995 khi p dng h thng thng k bnh nhn
hin i th s bnh nhn si ghi nhn c Anh v x Wales vo nm 1998 l
56 trng hp. en nm 2006 s bnh nhn mc loi bnh ny tng ln t bin:
tnh ring trong nm 2006 s bnh nhn si tng t 78 ti 740 trng hp,
nm 2007 l 990 trng hp v trong 9 thng u nm 2008 con s ny
tng ln ti 1049 trng hp.
Theo thng k ca T chc Y t Th gii (WHO), mi nm c khong 30
triu ngi trn ton Th gii b nhim si. Ch tnh ring nm 2006, c hn
240 ngn ngi, ch yu l tr em b t vong do mc si.
S d bnh si trong my nm tr li y tng t bin Anh v x Wales
l do ngy cng nhiu cc ng b, b m hoc l qun, hoc l t chi khng
tim vacxin phng bnh si, bnh quai b, bnh rubella cho tr.
M nguyn nhn ca vn ny l do vo nm 1998 c xut hin gi thit
v kh nng t min dch vi bnh si, bnh quai b, bnh rubella, do t nhiu
c nh hng n tm l ca cc bc cha m. Mc d khi , cc nh khoa hc
ln ting bc b gi thit ny, song t l min dch tr vn mc thp.
Ngi i din ca C quan Bo v sc kho cng ng ca Anh cho bit,
vic tr em khng tim phng bnh si, bnh quai b, bnh rubella c th l
nguyn nhn chnh dn ti s pht trin nhanh ca loi bnh ly nhim ny trong
thi gian gn y.

10

Cc chuyn gia d on rng, ch tnh ring Anh, s bnh nhn b bnh

si c th tng ln t 30.000 n 100.000 trng hp/nm. e ngn chn s ly
lan bnh truyn nhim ny sang cc khu vc khc cn phi tin hnh tim vacxin
m rng cho cc em hc sinh.
Bnh si cng l mt vn nghim trng ti Nht Bn. Ring
Hokkaido, trong s 541 trng hp mc bnh si, tr em chim khong 17,7%,
trong tr trn

thng tui chim

6 8 %,

95% s mc bnh u cha tim phng

vacxin. iu cho thy s cp bch ca vic tim phng cho tr ngay t khi
di

tui [16].

T vi chc nm nay, vi thuc chch nga si, bnh si t xy ra nhng

quc gia tin b, chng hn nh M, nhng vn cn l mt m nh kinh hong
cho nhng ngi dn cc nc ang pht trin.
Bnh li rt hay ly v d dng lan ra t nhng ngi du lch quc t. Hin
nay trn Th gii c n 30 ti 40 triu trng hp bnh si xy ra, gy ra hn
750.000 ci cht.
1.2.4.2. Tnh hnh mc bnh si Vit Nam
Hu nh khng mt ngi Vit Nam no khng tng nghe ni ti bnh
si. a b no b mc nhiu nt trong ngi cng c cho l ang b ban
si. Mt a b di 5 tui c th b rt nhiu bnh c km theo triu chng ni
nhng vt khp ngi v ln no cng cho l ln ban, phi king ra nng gi,
phi king nc, nu khng th s b chy hu.
T l mc bnh si ti Vit Nam gim r rt trong nhng nm qua, y
l hiu qu ca hot ng tim chng m rng (TCMR) c trin khai rng ri
trn c nc. Tuy nhin cho n nay bnh si vn cha c loi tr ti Vit
Nam. Thi tit sp chuyn sang ma ng xun - ma bnh si pht trin mnh

11

nn nhng hiu bit v cn bnh ny v cch phng chng s gip cng ng

hiu r hn gi tr ca vic phng bnh bng vacxin.
Chng trnh TCMR lm gim ng k t l t vong v mc bnh si
Vit Nam.T l lun hnh bnh si Vit Nam hin gim n 573 ln so vi
trc nm 1985, thi im bt u chng trnh TCMR. Cn bnh ny ang
c khng ch tt v d kin s b loi tr vo nm

2010.

Tnh hnh bnh si Vit Nam thi k trc khi tim 1 liu vacxin si
trong chng trnh TCMR cng tng t nh cc nc trn Th gii. Trong
thi k ny c ghi nhn min Bc Vit Nam vi t l mc bnh thp nht l
65/100.000 ngi dn (nm 1981), t l mc cao nht l 137,7/100.000 ngi
dn (nm 1979) v 125,7/100.000 ngi dn (nm 1983) v y l 2 nh ca
mt chu k dch si cch nhau khong t 4-5 nm. Bnh si lu hnh a
phng mi ni trong c nc v ph bin tr em, nht l tr di 5 tui.
Vic gy min dch phng bnh si bng mt liu vacxin si sng gim
c lc c bt u trong chng trnh TCMR Vit Nam t thng 10 nm
1985. Sau 5 nm, t l tim 1 liu vacxin si cho tr em t 9 - 11 thng tui
tng dn t 19% (nm 1985) ti 89% (nm 1989) v t l mc bnh si gim
xung t 91/100.000 ngi dn (nm 1986) xung 23/100.000 ngi dn (nm
1989). T n nm 2000, t l tim 1 liu vacxin si lun c duy tr trn
90% v t l mc bnh si gim n mc thp nht l 8,55/100.000 ngi dn
v cao nht l 19,01/100.000 ngi dn (nm 1993) v 23,16/100.000 dn (nm
2000) v y cng l 2 nh ca mt chu k dch si cch nhau khong t

-9

nm ca thi k sau khi tim vacxin si. Nh vy, sau 16 nm thc hin tim 1
liu vacxin si cho tr em t 9 - 11 thng tui t l mc bnh si gim xung
84%, chu k dch si ko di gp i v t l mc bnh cao c chuyn

12

dch ln nhm tui cao hn. Tuy nhin, bnh vn cn tn pht nhiu ni v vn
xy ra dch si vi quy m c nh hn thi k cha tim vacxin si. Tnh hnh
ny cng tng t nh cc nc khc trn Th gii.
Theo Cc Y t d phng (B Y t) cho bit, n nay c 30 tnh, thnh ph
c ngi b bnh si v st pht ban dng si; s ca dng tnh si l 650.
Ti H Ni ngy 23/02 c thm 29 ca st pht ban dng si, tng s t u
v dch n nay l 1.235 ca, trong 230 ca c xc nh dng tnh vi si,
hn 77% s ngi mc tui 18. Dch si xut hin ti 329 x, phng
trn tt c 29 qun, huyn ca H Ni. Theo Vin Cc bnh truyn nhim v
nhit i quc gia, t u v dch c 15 trng hp bin chng nng do si, hin
vn cn nm ca nng bin chng vim no, vim mng no.
Theo S Y t TP H Ch Minh, n ngy 23/02, thnh ph c 28 trng
hp mc bnh si.
Trung tm Y t d phng tnh H Tnh cho bit, sau na thng trin khai
phng, chng bnh si bn Ro Tre, huyn Hng Kh v huyn Nghi Xun,
tnh H Tnh. en nay, dch si c bn c khng ch, khng c trng hp
no mc thm. Dch si xut hin u tin vo ngy 05/02 bn Ro Tre. Ngi
bnh l H Th L (26 tui). Nhng nhng ngy sau c thm ba ngi bnh
khc ly bnh. V mt tun sau c 26 trng hp mc bnh, trong ngi
bnh hn 18 tui l 10 ngi. Cng trong thi gian ny, ti huyn Nghi Xun ri
rc c ba ngi bnh mc bnh si.
T ngy 03/02, trn a bn huyn Thanh H (Hi Dng) c hn 20
trng hp c biu hin st pht ban dng si, ch yu ti x Vit Hng, Tin
Tin, Thanh Hi. tui ca ngi b nhim bnh ch yu 18-40 tui.

13

T u thng 2 n nay, dch si pht trin nhanh ti nhiu a phng

trong tnh Nam nh, s liu ca S Y t cho bit, n ngy 24/02 pht
hin 27 bnh nhn st pht ban dng si, Hin c 9/10 huyn, thnh ph pht
hin ngi bnh mc dch si, gn y nht vo ngy 21/02 Trung tm Y t
d phng huyn V Bn pht hin thm mt trng hp c triu trng st
pht ban dng si.
Theo Trung tm Y t d phng tnh Ph Th, n nay ton tnh pht
hin 47 ngi bnh thuc 10/13 huyn, thnh ph, th x mc st pht ban dng
si Hu ht ngi bnh c tui t 14 n 30 [12].
Khi min dch gim th nguy c mc bnh s rt cao. l l do khin s
thanh nin mc bnh gia tng. Mt iu nguy him na l bnh si do virat ly
lan rt mnh, virut pht tn qua ng h hp vo khng kh, nhng ngi
khng c min dch hoc min dch km th kh nng b ly nhim cao.
1.3. Phng v iu tr bnh si
Si thng xut hin nhiu vo cui nm c, u nm mi. Khng ch gy
cc bin chng nh kit l ra mu mi, tiu chy ko di, vim phi, si cn c
th l nguyn nhn gy ci xng, suy dinh dng, thm ch t vong tr di 5
tui nu khng c iu tr ng.
1.3.1. Vacxin phng bnh si
Cch phng bnh tt nht l tim phng. Phng bnh bng vacxin c
khuyn co khi tr 9 thng tui, tuy nhin ngi ta thy rng vic tim mt mi
vacxin duy nht khng to ra min dch bn vng v rng ri trong cng ng
v t l tr tim phng bnh st cng nh t l t c min dch ca vacxin ny
cng ch t khong 90%. Do vy cn phi tim nhc li mi th 2, thi gian tim
l khi tr

tui, tui bc vo lp 1. Vic tim liu th 2 c th to min

14

dch ti 99%. Chng trnh TCMR Vit Nam thc hin mi nhc li ny trn
c nc t nm 2006. Ti cc a phng cn xut hin nhng v dch si nh,
i tng mc khng ch tr em th cn thit phi thc hin nhng chin dch tim
nhc cho ngi dn khu vc ny to min dch ln v bn vng trong cng
ng trong nhiu nm. C hai loi vacxin phng bnh si l vacxin bt hot bi
focmalin v vacxin sng gim c lc.
1.3.1.1. Vacxn bt hot
Vacxin bt hot bang formalin c ngun gc t chng Edmonston c
cp php ti M nm 1963 v c s dng cho n nm 1967.
Phc tim chng l 3 liu vacxin bt hot hoc hai liu vacxin bt hot
v mt liu vacxin sng c tim cch thng. Vacxin bt hot gy p ng
min dch ngn v c mt s phn ng ph c bit l mc si khng in hnh
nn sau t c s dng [16].
1.3.1.2. Vacxn sng gim c lc
Chng virut dng trong sn xut vacxin sng gim c lc c to ra
bng cch cy truyn lin tip trn cc t bo nui cy chn dng virut mt
c tnh nhng vn d tnh khng nguyn.
Nhiu loi vacxin gim c lc pht trin v c s dng c hiu qu
trn Th gii, bao gm cc chng Edmonston A v B do Enders phn lp c
dng rng ri nht hin nay. Vacxin ch t chng Moraten c s dng duy
nht ti M. Vacxin ch t chng Schwarz l sn phm chim u th ti nhiu
quc gia khc, ngoi ra mt s vacxin c ch t mt s chng khc gim c
nhiu hn nh AIK-C, Schwarz-F 8 8 , CAM-70, DT79 ang v c s dng
ti Nht [17].

15

Cng nh hu ht cc vacxin khc vacxin c sn xut t chng AIK-C

c sn xut trong t bo phi g nguyn pht. Loi vacxin ny bt u c
sn xut ti Vit Nam, B Y t giao cho Trung tm Nghin cu sn xut
vacxin v sinh phm y t (POLYVAC) thc hin d n sn xut vacxin si ti
Vit Nam. D n c thc hin bng ngun vn vin tr khng hon li ca
chnh ph Nht Bn. Mc tiu ca d n l xy dng mt c s sn xut vacxin
si vi cng sut 7,5 triu liu/nm t tiu chun GMP ca WHO. Cng ngh
sn xut vacxin do vin Kitasato, Nht Bn chuyn giao v chng AIK-C c
la chn sn xut vacxin [13].

Hnh 1: Vacxin si sn xut ti Vit Nam

1.3.1.3. Vacxin si kt hp vi vacxin quai b v rubella (MMR)
Cng nh nhiu nc trn Th gii, vic tim chng phng bnh si
thng c kt hp vi vacxin quai b v rubella ang c ngi dn Vit
Nam rt a chung. Vacxin hn hp ny c cp php nm 1971. Mt liu
MMR (0,5ml) cha t nht 1000 CCID 50 ca chng si Moraten, t nht 5000
CCID 50 chng quai b Jeryl Lynn v t nht

1000

CCID 50 chng virut vacxin

rubella RA27/3. C th c vacxin si phi hp vi rubella m khng c thnh

phn quai b. Hai loi vacxin MMR khc cng c cp php, mt loi c cha
chng si CAM-70 v loi kia cha chng Shawrz F 8 8 . s liu nghin cu min

16

dch v an ton cho thy vic kt hp khng nguyn si v khng nguyn quai
b, rubella rt an ton v hiu qu [17].
1.3.2. iu tr bnh si
Hin nay, chng ta vn cha c thuc iu tr c hiu i vi virut si. V
vy, vic chn on bnh sm c t ra nhm ngn chn s ly lan v pht
trin thnh dch nhng ni ng dn c.
Bnh nhn si c iu tr khng c hiu: ch dinh dng, ngh ngi
v iu tr triu chng:
+ Cho bnh nhn dng nhng thc n giu cht dinh dng, d tiu ha,
khng cn thit king c. Lu dng thm thc n nhiu vitamin A trnh
bin chng lot gic mc, m mt.
+ V sinh rng ming, da, mt.
+ H st: Lau mt, thuc h st thng thng (Paracetamol, khng dng
Aspirin v d gy hi chng Reyes).
+ Gim ho bang Dextromethorphan, Codein, thuc long m.
+ Khng Histamin: Pipolphen, Dimedron.
+ St trng mi hng, nh mt bng nc mui sinh l, vitamin A chng
kh mt.
+ Ch s dng khng sinh khi c bin chng: vim thanh qun, vim phi.
+ Cc bin php hi sc cp cu c p dng ty theo tnh trng ca
bnh nhn: th oxy h tr, th my, hi sc tim mch. [ 1 0 ]
1.4. Tnh hnh tim chng vacxin si phng bnh ti Vit nam

17

Tt c tr em Vit nam t 9 - 12 thng u c tim chng vacxin si

phng bnh si. Vacxin si l trong cc vacxin nm trong chng trnh Tim
chng m rng, tim min ph cho tr.
T trc ti nay vacxin si c khuyn co cn tim 1 mi, tuy nhin
trong thi gian gn y tnh hnh mc Si kh phc tp, do vy ch trng ca
Chng trnh tim chng m rng ang thc hin th im tim b sung mi 2
cho tr di

tui.

Liu lng : 0,5 ml/liu, trong cha > 3000 virut si/0,5 ml/liu
1.5. Sn xut vacxin si ti Vit nam
Mi nm chng trnh Tim chng m rng cn khong 1,5 triu liu
vacxin si cho tr trong tui 9 - 1 2 thng tui (tim mi 1 ) v tim b sung
mi

trong trng hp cn thit.

Vacxin si s dng cho chng trnh Tim chng m rng hin nay u l

vacxin nhp ngoi, l do l cha c vacxin si sn xut ti Vit nam.

Nm 2004, Vit Nam nhn c d n xy dng c s sn xut vacxin si
do Chnh ph Nht v t chc JICA gip bng ngun vn ODA khng hon
li. Polyvac l n v c B Y t ch nh thc hin d n ny. en thng
4/2006 nh my xy dng hon thnh, bt u giai on chuyn giao cng ngh
sn xut vacxin si. Thng 11/2007, 3 lot vacxin u tin sn xut thnh cng,
c cp chng ch cht lng v c php s dng tim th nghim trn
ngi tnh nguyn (th nghim lm sng).
Chng sn xut vacxin si l chng AIK-C, c sn xut trn trng g
sch nhp khu t c. Vacxin si sn xut ti Polyvac l vacxin sng gim c
lc ng kh.

18

1.6. Kim nh vacxin si sn xut ti Vit nam

Vacxin si sn xut ti Polyvac c cht lng theo tiu chun ca T chc
Y t Th gii (WHO) quy nh, sn phm thnh phm sau khi ng kh s c
kim tra cht lng thng qua 6 th nghim sau :
1) Th nghim v trng
2) Th nghim nhn dng
3) Th nghim hiu gi v n nh nhit
4) Th nghim m tn d
5) Th nghim an ton chung trn ng vt
6) Th nghim quan st trng thi vacxin
Th nghim hiu gi ca vacxin si l th nghim nhm xc nh hm
lng virut si. C nhiu phng php chun hiu gi nh :
- Chun vi lng, gy nhim trn t bo cm th Vero kn 1 lp, quan
st m hoi t v tnh hm lng virut theo cng thc Kaber hoc Reed Muench, n v tnh CCID 50/0,5 ml (liu hu hoi 50% t bo).
- Phng php to m hoi t, gy nhim trn t bo cm th Vero kn 1
lp, nhum v m s lng m hoi t, tnh hm lng virat, n v tnh
PFU/0,5 ml.
Hin ti Polyvac ang s dng phng php to m hoi t chun
hiu gi vacxin si sn xut ti c s.
Theo tiu chun ca WHO, vacxin si phi cha > 3000 CCID 50 / liu /
0,5 ml (hoc > 3000 PFU / liu). Tng ng > 3 lg CCID 50 / liu / 0,5 ml
(hoc > 3 lg PFU / liu)
1.7. Th nghim lm sng

19

Theo quy ch cp php s dng vacxin ca B Y t, mi vacxin mi u

phi qua giai on th nghim lm sng, l giai on th nghim trn ngi tnh
nguyn nhm nh gi: tnh an ton v tnh sinh min dch nhm bo v ngi
tim vacxin khng mc bnh si.
Vacxin si sn xut ti Polyvac mc d c quy trnh cng ngh ging vi
vacxin ang sn xut ti Nht Bn, nhng cha tng c s dng ti Vit Nam
nn cng c coi nh l vacxin mi, bt buc phi thc hin th nghim lm
sng trn ngi tnh nguyn.
Tt c tr tnh nguyn u c kim tra sc khe v ly mu kim tra
khng th si trc khi tim, sau tim 6 tun ly mu kim tra khng th si,
nh gi mc p ng min dch.

20

Chng 2 - VT LT V PHNG PHP NGHIN c u

2.1. Phng php chun hiu gi bng to m hoi t
2.1.1. Nguyn vt liu
2.1.1.1. Mu chun
S dng mu chun vacxin si do Vin Kitasato Nht Bn cung cp, k hiu
mu chun M I6-6. Hiu gi qun l 104,2 - 1046 PFU/0,5ml ). Bo qun - 70c
2.1.1.2. Ho cht, mi trng
1. Nc hi chnh vacxin

(Polyvac sn xut)

2. MEM

(Polyvac pha)

3. 7,5% N aH C03

(Polyvac pha)

4.

Khng sinh P-S

(Polyvac pha)

5. Huyt thanh b (BS)

(Gibco - M)

6. Glutamin 200mM

(Polyvac pha)

7. Huyt thanh b bo thai (FBS) (Code No. 16140-071, Gibco - M)

8. Agarose ME

(Code No. 50013R-Iwai - Nht)

9. EagleMEM

(Code No. 05901-N issui)

10. trung tnh 0,2% (Neutral Red )

(Polyvac pha)

2.1.1.3. Dng c
Dng c s dng phi c tit trng m bo v trng. Phng php tit
trng s dng l : sy kh, hp t hoc chiu tia ...
1. Pipet lOml

(Code No.7087-10, Pyrex - M)

2. Pipet Aid

(Code No. 4910, Coning - M)

3. Pipet man 10 ~ 200|il

(Code No. F 123601, Gilson - Php)

4.

Pipet Man

5. Pipet Man

100 ~ lOOOLil (Code No. F123602, Gilson - Php)

lml ~ 5ml

6. u tp 10 ~ 200^1

(Code No. F123603,Gilson - Php)

(Coning - M)

21

7. u tp 100 ~ 1000jil

(Coning - M)

8. u tp lml ~ 5ml

(Coning - M)

9. Tp thu tinh y trn 5ml

(Code No. 9820-12, Pyrex - M)

10. Chai thu tinh 1OOml

(Dural - c)

11. Chai thu tinh 1OOOml

(Dural - c)

12. Gi cm tp

(Vit Nam )

13. Bm tim chia liu 5- lOml (Socorex- Nht)

14. My trn

(Code.No TM- 1 ,AS ONE )

15. Knh hin vi

(Code No. 1X51- 71 PHP,OLYMPUS )

16. T m C 0 2

(Code No. MC0175 , SANYO

17. B iu nhit

(AS ONE)

18. Khn khng sinh bi

(Hn Quc)

19. Bnh ng cha mi trng thi (Code No. 2126-5000, Nalgene- M)

20. Phu inox

(Vit Nam)

21. Phin nui t bo 6 ging

(Coning - M)

22. Nhn ghi tn mu

(Nht)

23. Chai 2,5L

(Trung Quc)

24. Bt pht

(Nht)

25. Chu nha

(Vit Nam)

2.1.1.4. Te bo s dng
Te bo Vero (FV) l t bo cm th vi virat si c s dng chun hiu
gi vacxin si. T bo Vero c nui trn phin 6 ging vi nng 20xl04t bo /
ml X 3ml / ging. Nui 37c lc , 5% C 2 trong vng 2 - 3 ngy, t bo kn 1 lp
dng chun .
2.1.1.5. Mi trng s dng
1. GM5%BS mi trng pht trin nui t bo v pha long mu

22

2. Mi trng ph thch ln 1: Hn MEM 2 ln (khng c trung tnh) vi

Agarose 1% theo t l 1:1
3. Mi trng ph thch ln 2 : Hn MEM 2 ln (c trung tnh) vi Agarose 1%
theo t l 1:1
Chun bi MEM 2 ln
-

Pha MEM 3 ln (1000 ml)

+ Eagle MEM

28,2g

+ WFI va

1000ml

Sy tit trng 1210C / 15 pht

Chia ra c h a i: 66,7ml / chai (s dng chai sy tit trng)
-

S dng FBS bt hot pha MEM 2 ln

S dng nc WFI sy tit trng 1210C/30 pht pha MEM 2 ln

MEM 2 ln
(Khng c trung tnh) (1) (ml)
66,7

MEM 2 ln
(C trung tnh) (2) (ml)
66,7

FBS

4,0

4,0

7,5% NaHCOs

4,0

4,0

Glutamin 200 mM

2,0

2,0

p-s

trung tnh 0,2%

10

22,3

12,3

100

100

Thnh phn
MEM 3 ln

WFI
m A

Tng

MEM 2 ln ngm b iu nhit nhit ~ 44

trn vi Agarose 1% ph thch.
Chun bi pha Agarose 1%
+ Agarose
+ WFI va

1,0 g
100 ml
23

c khong 1 0 - 1 5 pht trc khi

121c/15 pht

sy tit trng

Agarose 1% ch pha s dng trong ngy.

Ch s dng chai thy tinh loi 500 ml pha thch Agarose 1%
Sau sy tit trng, ly ra khi l sy bo qun nhit ~

44c trong b iu

nhit cho n khi trn vi MEM 2 ln.

Pha mi trng GM5% BS
Thnh phn

Th tch (ml)

MEM

930

BS

50

NaHCOs 7,5%

15

Khng sinh (P- S)

Tng
2,1,2, Quy trnh thc hin

1000

2.1.2.1. Chun b mu v mu chun

1. Chun b hp xp mu c sn vn
2. Ly mu v mu chun t t ct mu ra t vo hp xp chun b trn
3. Vn chuyn hp cha mu n ni th nghim. Mu c bo qun nh vy
cho n khi pha long
2.1.2.2. Pha long, gy nhim, nui
Thc hin trong iu kin v trng, trong Clean bench
1.

Chun b cho pha long mu v mu chun bc 5 v 10: xp tp pha long ra

gi cm tp, cn c vo s lng mu v s nng pha long xp theo th t. Ht

l,8ml GM5%BS cho tp pha long bc 10 (1 lg) v ht l,lml GM5%BS cho tp pha
long bc 5 (0,5 lg )

24

2. Ly cc khay t bo (phin 6 ging) t t 37c, 5%C2 ra, soi trn knh hin vi
quan st t bo, kim tra tnh trng t bo. Te bo kn u 1 lp iu kin cho th
nghim.
3. Loi b nc ni nui t bo trong phin (s dng chai nha 5L v phu inox
thc hin thao tc ny) tri khn khng sinh bi thm cn ln bn. p ngc phin ln
khn khng sinh bi thm cn chun b sn thm ht mi trng cn st li trong
ging t bo.
4. Dn nhn v pha bn phi phin vi thng tin: tn mu, s phin
5. Hi chnh vacxin: 0,7 ml (i vi mu chun M I6-6) v 5,5 ml (i vi vacxin
mu do Polyvac sn xut)
6. Pha long bc 10: cho 0,2ml mu vo tp c sn l,8ml GM5% BS > trn u,
(mu c pha long nng 101 tp ny). Pha long nng tip theo, thm
0,2ml mu nng 10'1 vo tp cha l,8ml mi trng tp k tip > trn u,
c mu pha long 10"2 tip tc lm nh vy n khi ht nng pha long
7. Pha long bc 5: cho 0,5 ml mu pha long 1CT1 sang tp cha 1,1 ml GM5%BS
*trn u, mu c pha long 10"1 . Pha long 10"2,5 cng thc hin nh vy t tp
pha long 102 tip tc lm nh vy n khi ht nng pha long
V d : Quy trnh pha long nh sau:
Bc pha long

101

Mu

0,2 ml

GM5%BS

1,8 ml

Bc pha long

10 2
1

0,2 ml "1
1,8 ml
1 0 -1,5

10 3
0,2 ml
1,8 ml
! 0-2,5

104
0,2 ml
1,8 ml
1 0 -3,5

Mu

0,5 ml

0,5 ml

0,5 ml

GM5%BS

1,1 ml

1,1 ml

1,1 ml

8. 3 nng pha long cui cng s c gy nhim, gy nhim 0,lml/ging x2

ging/1 nng . Di y l s minh ho vic gy nhim trn phin 6 ging

25

10'3

103'5 10'4

9. Lp li bc (6) v (7) vi cc mu v mu chun cn li cho n ht

10. Sau gy nhim lng phin sang tri, phi lng mu gy nhim ph kn b
mt t bo, xp phin vo khay hp ph iu kin 37 lc, 5%C2 trong 60 pht
11. Ket thc thi gian hp ph ly khay t bo t vo Clean bench, ly MEM 2 ln
v Agarose 1% chun b sn t b iu nhit ra, dng khn khng sinh bi thm cn
lau sch bn ngoi 2 chai v chuyn vo Clean bench, lau li mt ln na trong clean
bench. chai MEM 2 ln vo chai Agarose (t l 1:1) lc u, dng pipet thu tinh
lOml v trng ph thch, ph 3ml/ging. Ph xong phin no phi lc nh phin
cho u mi trng gy nhim v mi trng ph thch ho u vo nhau.
12. e nguyn s phin va ph thch ti ch cho n khi ng hon ton (20 ~ 30
pht) nui iu kin 37 lc, 5% C02 trong 7 ngy
13. Sau 7 ngy nui cy iu kin 37 lc, 5%C2 tin hnh ph thch ln 2 c
trung tnh, thnh phn ca ph thch ln 2 nh m t 2.1.1.5
14. Tip tc nui 37 lc, 5% C02 trong vng 3 - 4 ngy
15. m PFU (mu trng trn nn mu )
16. Tnh kt qu
2.1.2.3.

Phng php tnh kt qu

Cn c vo s lng m hu hoi m c (PFU)

Quy tc tnh kt qu
1. Tnh hiu gi ti tng nng pha long,

26

2. Tnh gi tr trung bnh gia cc nng pha longca cng 1 mu

3. Tnh gi tr trung bnh ca mu
4. Chuyn i n v tnh hiu gi
Ch tnh kt qu vi cc iu kin sau
a. Nng gy nhim c s m PFUtrong khong > 10 v <100 PFU c
2 phin gy nhim
b. Cng nng gy nhim c s m PFU chnh nhau <3,16 ln (tng
ng 0,5 lg PFU/0,5 ml)
c. Cng mt mu c 2/3 nng gy nhim tho mn iu kin a v b
V d tnh nh sau
10'2 10'2,5 10'3

Tnh hiu gi ti cc nng gy nhim

Ti nng gy nhim 102
M = ( 55 + 52 ) / 2 = 53,5
Hiu

gi = lgio2+ lg 53,5 =2 + 1,73 = 3,73 (1)

Ti nng gy nhim 102,5

M = ( 26 + 21 )/2 = 23,5
Hiu gi = lg 102,5 + lg 23,5 = 2,5 + 1,37 = 3,87 (2)
Tnh hiu gi ca mu
Hiu gi mu = [ (1) + (2) ] /2 = ( 3,73 + 3,87 ) / 2 = 3,8 PFU / 0,lm l (3)
Chuyn i : (3) + lg5 = 3,8 + 0,7 = 4,5 PFU / 0,5ml

27

Tnh hiu gi trung bnh

Hiu gi mu

(1) = 1o4,50

Hiu gi mu (2) = 1o4,44 >Hiu gi mu (3) = 1o4,52

^4,49

> Trung bnh = 10

Ket qu chun hiu gi c gi tr khi tha mn 2 iu kin sau :

1) Hiu gi mu chun M I6-6 (Vin Kitasato - Nht Bn cung cp) nm trong
khong 4,2 ~ 4,6 lg PFU / 0,5ml.
2) Chnh lch hiu gi gia cc l mu ca cng mt mu trong cng mt ln
th nghim < 0,5 lg PFU / 0,5 ml
2.2. Phng php kho st hiu gi vacxn si dng cho th nghim lm sng
2.2.1. Phng php ly mu
3

lot vacxin lin tip sn xut 11/2007 c tim th nghim lm sng trn tr

em tnh nguyn ti huyn An Lo - Hi Phng. Qu trnh thc hin th nghim lm

sng t 3 -7/11/2008.
M s 3 lot vacxin : M-0107 ; M-0207 v M-0307
kt qu th nghim lm sng c tnh khch quan, chng ti la chn vacxin
Rouvac hin ang c s dng trong chng trnh Tim chng m rng tim i
chiu, vacxin i chiu c k hiu R.
Trong khun kh ca kha lun chng ti chn ngu nhin 5 x ly mu kho
st hiu gi trc tim v hiu gi thu hi sau tim. 5 x gm: Bt Trng, An Thi,
Quang Trung, Trng Sn v An Tin.
Mi x u tim 3 loi vacxin do Polyvac sn xut v vacxin i chiu.

28

Mu vacxin trc tim: ly ngay ti kho ca Polyvac trc khi giao mu cho
nhm vn chuyn mu n a im tim th nghim lm sng. Mu c bo qun <

8c (thng thng bo qun -30c ti Polyvac) cho n khi kim tra hiu gi. Nh vy
trc tim c 4 mu: M -0107(T); M -0207(T); M-0307(T) v R(T)
Mu thu hi sau tim: thu hi l vacxin ngay ti bn tim ca 5 x (l nguyn
cha m np, c bo qun trong hp xp c phin lnh trong sut thi gian tim),
vn chuyn v Polyvac cng trong iu kin bo qun lnh (<8C) cho n khi kim
tra hiu gi. Nh vy mu thu hi sau tim l 20 mu (4 mu/1 x), k hiu mu: M0107/Bt Trng,...
2.2.2. Quy trnh chun hiu gi
Thc hin chun hiu gi 24 mu trn theo quy trnh m t 2.1.2
2.2.3. Phng php kho st
Tiu chun: > 1000 PFU / 0,5 ml / 1 liu, tng ng > 3 lg PFU / 0,5 ml / 1
liu (theo tiu chun ca WHO)
Kho st kt qu 4 mu vacxin trc tim v 20 mu thu hi sau tim.

29

Chng 3 - KT QU V BN LUN
3.1. Kt qu xc nh hiu gi vacxin si mu M-0107
Bng 1: Ket qu xc nh hiu gi vacxin si mu M-0107
Tn M u
Trc tim
M-0107(T)
M -0107/Bt
Trng
(Sau tim)
M-0107/An
Thi
(Sau tim)
M-0107/uang
Trung
(Sau tim)
M-0107/Trng
Sn
(Sau tim)
M-0107/An
Tin
(Sau tim)
MI 6-6
(Mu chun)

Nng gy nhim
1 0 -2,5

1C-2
71
55
57
73
52
48
46
50
47
67
65
59
66
50
67
102
96
76
41
38
60

65
70
68
62
56
66
40
45
40
80
72
63
72
49
52
95
75
78
35
49
43

19
22
15
19
13
19
22
18
17
24
20
19
22
18
14
38
19
29
16
16
25

1Cr 3
4
10
4
8
4
6
8
8
6
6
6
7
6
7
3
8
13
2
6
6
3

19
15
19
24
20
14
11
30
25
17
17
19
15
20
15
35
20
19
11
18
21

11
6
6
6
3
9
7
5
7
5
6
7
5
3
3
9
10
7
6
4
2

Hiu gi
lgPFU/0,5ml
4,51
4,59
4,47
4,53
4,43
4,44
4,38
4,48
4,43
4,54
4,51
4,49
4,51
4,44
4,38
4,76
4,63
4,59
4,31
4,39
4,49

Hiu gi
tru n g bnh
4,52

4,47

4,43

4,51

4,44

4,66

4,40

T kt qu bng 1 cho thy tt c cc mu, s lng m hoi t thu c

u tun theo quy tc chun hiu gi, iu kin tnh kt qu (xem mc a, b v c
trang 28).
Hiu gi ca 3 l mu ca cng 1 mu u khng chch lch qu 0,5 lg PF / 0,5
ml. C th: M-0107 (T) c hiu gi ln lt l 4,51; 4,59 v 4,47. M-0107/Bt Trng c
hiu gi ln lt l 4,53; 4,43 v 4,44. M-0107/An Thi c hiu gi ln lt l 4,38; 4,48
v 4,43. M-0107/Quang Trung c hiu gi ln lt l 4,54; 4,51, v 4,49. M-

30

0107/Trng Sn c hiu gi ln lt l 4,51; 4,44 v 4,38. M-0107/An Tin c hiu gi

ln lt l 4,76; 4,63 v 4,59. M I6-6 c hiu gi ln lt l 4,31; 4,39 v 4,49.
Mu vacxin chun M I6-6 c hiu gi trung bnh: 4,40 lg PFU / 0,5 ml, nm
trong khong hiu gi qun l (4,2 ~ 4,6 lg PFU / 0,5 ml).
Tt c 5 mu vacxin thu hi ti bn tim (sau tim) ca 5 x v mu vacxin
trc tim (ca lot M-0107) u > 3 lg PFU / 0,5 ml, tha mn tiu chun ca WHO.

KHO ST HIU GI VACXIN S (M-0107) TT CC X

lg PFU/ 0,5 ml

M -0107(T )

M -0107/B t

M -0107/A n

M -0107/ uang

M-

T rang

Thi

T rung

0107/Trng

M -0107/A n

M 16-6

Tin

Sn

Hiu gi

----- Tiu chun

------ Thp

------ Cao

th 1: S phn b hiu gi mu M-0107 trc tim v ti cc x

Nhn trn th 1 thy vacxin M-0107 trc khi vn chuyn n a im tim
c hiu gi trung bnh 4,52 lg PFU / 0,5 ml. Sau khi vn chuyn ti 5 x tim

cho

tr tnh nguyn thu hi v hiu gi trung bnh giao ng t 4,44 - 4, lg PFU /0,5 ml.
Mu vacxin c hiu gi thp nht (4,44) thu hi t x Trng Sn, thp hn mu trc
khi giao cho nhm th nghim lm sng (< 0,08 lg PFU / 0,5 ml), tuy nhin vn t
31

tiu chun v hiu gi (> 3 lg PF / 0,5 ml). Mu vacxin c hiu gi trung bnh cao
nht (4,66) thu hi t x An Tin, cao hn c mu trc khi giao cho nhm tim th
nghim lm sng (> 0,14 lg PF / 0,5 ml). Chnh lch hiu gi ny < 0,5 lg PF / 0,5
ml, sai s cho php trong qu trnh chun hiu gi cc vacxin virut.
3.2. Kt qu xc nh hiu gi vacxin si mu M-0207
Bng 2: Ket qu xc nh hiu gi vacxin si mu M-0207
Nng gy nhim

Tn M u
Trc tim
M-0207(T)
M-0207/Bt
Trng
(Sau tim)
M-0207/An
Thi
(Sau tim)
M-0207/Quang
Trung
(Sau tim)
M-0207/Trng
Sn
(Sau tim)
M-0207/An
Tin
(Sau tim)
MI 6-6
(Mu chun)

1 0 -2,5

10'2
59
58
47
44
51
37
52
59
50
56
66
43
44
64
58
105
60
74
31
43
22

51
65
49
40
59
33
60
50
57
80
97
73
62
53
48
106
74
77
38
29
46

20
14
9
16
14
13
17
18
12
19
19
25
26
22
26
30
19
22
11
10
15

1o 3
14
17
12
10
24
12
18
15
14
33
24
11
17
21
14
35
27
35
16
16
10

3
10
4
5
3
9
2
7
2
12
7
2
11
10
3
8
10
7
7
3
2

9
6
8
4
7
2
7
8
5
10
12
8
9
9
9
8
6
9
2
2
1

Hiu gi
lgPFU/0,5ml
4,44
4,44
4,38
4,32
4,46
4,27
4,42
4,43
4,37
4,63
5,57
4,51
4,48
4,50
4,46
4,71
4,55
4,62
4,29
4,29
4,27

Hiu gi
tru n g bnh
4,42

4,35

4,41

4,57

4,48

4,63

4,28

T kt qu bng 2 cho thy tt c cc mu, s lng m hoi t thu c u

tun theo quy tc chun hiu gi, iu kin tnh kt qu (xem mc a, b v c trang 28).
Hiu gi ca 3 l mu ca cng 1 mu u khng chch lch qu 0,5 lg PFU / 0,5
ml. C th: M-0207(T) c hiu gi ln lt l 4,44; 4,44 v 4,38. M-0207/Bt Trng c
hiu gi ln lt l 4,32; 4,46 v 4,27. M-0207/An Thi c hiu gi ln lt l 4,42; 4,43
32

v 4,37. M-0207/Quang Trung c hiu gi ln lt l 4,63 ; 4,57 v 4,51. M0207/Trng Sn c hiu gi ln lt l 4,48; 4,50 v 4,46. M-0207/An Tin c hiu gi
ln lt l 4,71; 4,55 v 4,62. M I6-6 c hiu gi ln lt l 4,29; 4,29 v 4,27.
Mu vacxin chun M I6-6 c hiu gi trung bnh: 4,28 lg PFU / 0,5 ml, nm
trong khong hiu gi qun l (4,2 ~ 4,6 lg PFU / 0,5 ml).
Tt c 5 mu vacxin thu hi ti bn tim (sau tim) ca 5 x v mu vacxin
trc tim (ca lot M-0207) u > 3 lg PFU / 0,5 ml, tha mn tiu chun ca WHO.
K H O S T H I U G I VA CX IN S I (M -0207) T I C C X
lg PFU / 0,5 ml

5
4,57

4.5

4,42

-M5-

4.41

4,63
4.48

J,6_
4,28

4,2

4
3.5
3
2.5
M-0207(T)

M-0207/Bt
Trang
Hiu gi

M-0207/An Thi M-0207/uang

Trung
Tiu chun

M-0207/Tnrng M-0207/An Tin

Sn
Thp

M16-6

Mu

Cao

th 2: S phn b hiu gi mu M-0207 trc tim v ti cc x

Nhn trn th 2 thy vacxin M-0207 trc khi vn chuyn n a im tim
c hiu gi trung bnh 4,42 lg PFU / 0,5 ml. Sau khi vn chuyn ti 5 x tim cho
tr tnh nguyn thu hi v hiu gi trung bnh giao ng t 4,35 - 4,63 lg PF /0,5 ml.
Mu vacxin c hiu gi thp nht (4,35) thu hi t x Bt Trng, thp hn mu trc
khi giao cho nhm tim th nghim lm sng (< 0,07 lg PF / 0,5 ml), tuy nhin vn
t tiu chun v hiu gi (> 3 lg PFU / 0,5 ml). Mu vacxin c hiu gi trung bnh cao

33

nht (4,63) thu hi t x An Tin, cao hn c mu trc khi giao cho nhm tim th
nghim lm sng (> 0,21 lg PF / 0,5 ml). Chnh lch hiu gi ny < 0,5 lg PF / 0,5
ml, sai s cho php trong qu trnh chun hiu gi cc vacxin virat.
3.3 Kt qu xc nh hiu gi vacxin si mu M-0307
Bng 3 : Ket qu xc nh hiu gi vacxin si mu M-0307
Tn M u
Trc tim
M-0307(T)
M-0307/Bt
Trng
(Sau tim)
M-0307/An Thi
(Sau tim)
M-0307/Quang
Trung
(Sau tim)
M-0307/Trng
Sn
(Sau tim)
M-0307/An Tin
(Sau tim)
M I 6-6
(Mau chun)

10'2
61
64
64
61
47
46
59
82
54
68
89
63
80
65
57
111
90
98
51
62
57

Nng gy nhim
10 25
84
22
26
12
21
63
20
21
59
53
17
23
15
10
49
15
46
18
14
57
15
92
21
20
54
14
10
86
30
31
27
87
37
55
18
20
75
26
20
21
86
28
69
19
20
112
30
32
85
22
19
94
25
16
14
17
66
14
51
16
45
16
16

1o 3
9
4
6
8
5
2
4
8
5
14
7
11
6
6
15
19
10
12
4
4
5

8
9
9
12
5
8
7
5
6
14
10
4
5
9
6
13
10
9
2
2
2

Hiu gi
lgPFU/0,5ml
4,57
4,46
4,50
4,48
4,34
4,39
4,41
4,58
4,36
4,71
4,68
4,48
4,58
4,59
4,50
4,80
4,62
4,60
4,44
4,42
4,40

Hiu gi
tru n g bnh
4,51

4,40

4,45

4,62

4,56

4,67

4,42

T kt qu bng 3 cho thy tt c cc mu, s lng m hoi t thu c

u tun theo quy tc chun hiu gi, iu kin tnh kt qu (xem mc a, b v c
trang 28).
Hiu gi ca 3 l mu ca cng 1 mu u khng chch lch qu 0,5 lg PFU / 0,5
ml. C th: M-0307(T) c hiu gi ln lt l 4,57; 4,46 v 4,50. M-0307/Bt Trng c

34

hiu gi ln lt l 4,48; 4,34 v 4,39. M-0307/An Thi c hiu gi ln lt l 4,41; 4,58

v 4,36. M-0307/Quang Trung c hiu gi ln lt l 4,71; 4,68 v 4,48. M0307/Trng Sn c hiu gi ln lt l 4,58; 4,59 v 4,50. M-0307/An Tin c hiu gi
ln lt l 4,80; 4,62 v 4,60. M I6-6 c hiu gi ln lt l 4,44; 4,42 v 4,40.
Mu vacxin chun M I6-6 c hiu gi trung bnh: 4,42 lg PFU / 0,5 ml, nm
trong khong hiu gi qun l (4,2 ~ 4,6 lg PFU / 0,5 ml).
Tt c 5 mu vacxin thu hi ti bn tim (sau tim) ca 5 x v mu vacxin
trc tim (ca lot M-0307) u > 3 lg PFU / 0,5 ml, tha mn tiu chun ca WHO.

KHO ST HIU GI VACXIN SI (M-0307) TI CC X

lg PFU/ 0,5 ml

M -0307(T )

M -0307/Bt

M -0307/A n

M -0307/ uang

M-

M -0307/A n

Trang

Thi

T rung

0307/T rng

Tin

M 16-6

Sn

I Hiu gi

---------- Tiu chun

--------- Thp

------Cao

th 3: S phn b hiu gi mu M-0307 trc tim v ti cc x

Nhn trn d th 3 thy vacxin M-0307 trc khi vn chuyn dn a im tim
c hiu gi trung bnh 4,51

lg PFU / 0,5 ml. Sau khi vn chuyn ti 5 x tim cho

tr tnh nguyn thu hi v hiu gi trung bnh giao ng t 4,40 - 4,67 lg PFU /0,5 ml.
35

Mu vacxin c hiu gi thp nht (4,40) thu hi t x Bt Trng, thp hn mu trc

khi giao cho nhm tim th nghim lm sng (< 0,11 lg PFU / 0,5 ml), tuy nhin vn
t tiu chun v hiu gi (> 3 lg PFU / 0,5 ml). Mu vacxin c hiu gi trung bnh cao
nht (4,67) thu hi t x An Tin, cao hn c mu trc khi giao cho nhm tim th
nghim lm sng (>0,16 lg PFU / 0,5 ml). Chnh lch hiu gi ny < 0,5 lg PFU / 0,5
ml, sai s cho php trong qu trnh chun hiu gi cc vacxin virat.
3.4 Kt qu xc nh hiu gi vacxin si i chng (R)
Bng 4 : Kt qu xc nh hiu gi vacxin si i chng (R)
Tn M u
Trc tim
R(T)
R/Bt Trng
(Sau tim)
R/An Thi
(Sau tim)
R/Quang
Trung
(Sau tim)
R/Trng Sn
(Sau tim)
R/An Tin
(Sau tim)
MI 6-6
(Mu chun)

>100
78
>100
57
46
37
38
39
40
38
53
61
74
71
86
56
69
55
47
35
34

Nng gy nhim
0'2
10 25
34
>100
32
41
95
31
>100
38
38
11
10
49
63
17
21
44
13
10
13
12
45
48
15
21
60
10
16
65
16
22
22
17
49
24
25
69
24
98
30
55
20
19
28
32
87
20
13
68
20
69
19
63
15
19
36
10
13
12
11
50
12
14
49

1(r 3
14
10
8
4
4
1
5
5
4
4
2
10
8
10
7
2
6
6
5
2
5

11
9
7
4
3
7
8
8
3
5
6
9
7
9
10
6
8
4
4
5
5

Hiu gi
IgPF/0,5ml
4,76
4,70
4,78
4,32
4,46
4,29
4,31
4,40
4,36
4,45
4,45
4,55
4,63
4,50
4,66
4,46
4,52
4,45
4,29
4,30
4,32

Hiu gi
tru n g bnh
4,75

4,36

4,36

4,48

4,60

4,48

4,30

T kt qu ghi bng 4 cho thy tt c cc mu, s lng m hoi t thu

c u tun theo quy tc chun hiu gi, iu kin tnh kt qu (xem mc a, b
v c trang 28).
36

Hiu gi ca 3 l mu ca cng 1 mu u khng chch lch qu 0,5 lg PF / 0,5

ml. C th: R(T) c hiu gi ln lt l 4,76; 4,70 v 4,78. R/Bt Trng c hiu gi ln
lt l 4,32; 4,46 v 4,29. R/An Thi c hiu gi ln lt l 4,31; 4,40 v 4,36. R/Quang
Trung c hiu gi ln lt l 4,45; 4,45 v 4,55. R/Trng Sn c hiu gi ln lt l
4,63; 4,50 v 4,66. R/An Tin c hiu gi ln lt l 4,46; 4,52 v 4,45. M I6-6 c hiu
gi ln lt l 4,29; 4,30 v 4,32.
Mu vacxin chun M I6-6 c hiu gi trung bnh: 4,30 lg PFU / 0,5 ml, nm
trong khong hiu gi qun l (4,2 ~ 4,6 lg PFU / 0,5 ml).

lg P F U / 0,5 ml

KHO ST HIU GI VACXIN SI I CHIU (R) TI CC X

4,75
4,48

4,6

4^36---------------4^36

4,5

4,3

4,2

3,5

2,5
R(T)

R/tt Trang
I Hiu gi

R/An Thi

R/QuangTrung

Ticu chun

R/TrngSn
Thp

R/An Tin

M16-6

]V[u

Cao

th 4: S phn b hiu gi mu vacxin chun (R) trc tim v ti cc x

Nhn trn th 4 thy vacxin M-0407 trc khi vn chuyn n a im tim
c hiu gi trung bnh 4,75 lg PFU / 0,5 ml. Sau khi vn chuyn ti 5 x tim cho
tr tnh nguyn thu hi v hiu gi trung bnh giao ng t 4,36 - 4,60 lg PFU /0,5 ml.
Mu vacxin c hiu gi thp nht (4,36) thu hi t 2 x Bt Trng v An Thi, thp

37

hn mu trc khi giao cho nhm tim th nghim lm sng (< 0,39 lg PF / 0,5 ml),
tuy nhin vn t tiu chun v hiu gi (> 3 lg PF / 0,5 ml). Mu vacxin c hiu gi
trung bnh cao nht (4,60) thu hi t x Trng Sn cng vn thp hn mu trc khi
giao cho nhm tim th nghim lm sng (< 0,15 lg PFU / 0,5 ml). Chnh lch hiu
gi ny < 0,5 lg PFU / 0,5 ml, sai s cho php trong qu trnh chun hiu gi cc
vacxin virut.
Bng 5: Kho st bin ng hiu gi trong qu trnh th nghim lm sng

Trc tim
Bt Trng
An Thi
Quang Trung
Trng Sn
An Tin
Trung bnh
lch chun
Confidence

M-0107

M-0207

M-0307

4,52
4,47
4,43
4,51
4,44
4,66
4,51
0,08
4,51 0,07

4,42
4,35
4,41
4,57
4,48
4,63
4,48
0,11
4,48 0,08

4,51
4,40
4,45
4,62
4,56
4,67
4,54
0,10
4,54 0,08

R
(vacxin i chng)
4,75
4,36
4,36
4,48
4,60
4,48
4,51
0,15
4,51 0,12

Nh bit vacxin Si l vacxin yu d cht bi nh sng v nhit . Chnh v

vy vacxin phi bo qun di dng ng kh v trong thnh phn phi c cht bo
qun nh: gelatin, sacarose, sorbitol,...
Vit nam l mt nc nhit i, nhit thng xuyn cao, vic vn chuyn
vacxin ti a im tim phi lun c bo qun trong iu kin ti v mt (< 8C).
Trong kt qu kho st hiu gi vacxin cho thy c 3 lot vacxin sn xut ti
Polyvac cc thi im khc nhau ca qu trnh th nghim lm sng u t tiu
chun > 3 lg PFU / 0,5 ml v s thay i hiu gi trc tim v sau tim khng c
ngha thng k (bng 5), tng ng nhm vacxin i chng (vacxin ang c s
dng tim cho tr em Vit Nam). Nh vy vic bo qun vacxin trong hp xp c bnh
tch lnh bo qun tt vacxin si cho qu trnh tim th nghim lm sng.
38

KT LUN
1. Hiu gi ca tt c 3 lot vacxin sn xut ti Polyvac u > 1000 PFU / 0,5
ml, t tiu chun ca T chc Y t Th gii (WHO)
2. Hiu gi ca cc mu vacxin thu hi ti bn tim ca 5 x u > 1000 PFU /
0,5 ml, t tiu chun ca T chc Y t Th gii (WHO)
3. S bin ng v hiu gi ca vacxin do Polyvac sn xut trong qu trnh bo
qun, vn chuyn n a im tim tng ng vi vacxin i chng (vacxin ang
s dng trong chng trnh Tim chng m rng ti Vit Nam)

39

TT LTU THAM KHO

TING VIT
1. http://www.pasteur-hcm.org.vn/ytecongdong/tiemchung/dichtehoc-benhsoi
2. B Y t (2001), Tng kt cng tc tim chng m rng nm 2001, H Ni.
3. Nguyn nh Bng, Hong Ngc Hin, Phm L Hng, m Vit Cng
(1992), Vi sinh vt Y hc, Nh xut bn i hc v Trung hc chuyn nghip,
H Ni.
4. Bi i (1982), Bnh hc truyn nhim, Hc vin Qun y, H Ni, tr 121-130.
5. Hunh Phng Lin, Nguyn Kim Tho (1991), ATv thut xt nghim vi sinh vt
y hc, Nh xut bn vn ho, H Ni.
6. Jdanov VM.,

s. Ya Gaidamovich (1979), Virut hc (on Xun Mu dch),

Nh xut bn Y hc, H Ni
7. Nguyn Hnh Phc (1991), Tnh ch khng nguyn vrut si ng dng trong cc
k thut chun on huyt thanh hc bnh si, Lun n Ph tin s khoa hc Y
Dc, Vin V sinh dch t hc, B Y t, H Ni.
8. Nguyn Hnh Phc, Nguyn Vn Trang (2000), Chn on huyt thanh hc
bnh si, Tuyn tp cng trnh 1997-2000, Vin V sinh dch t Trung ng,
Nh xut bn Y hc H Ni, tr 20-25.
9. Phm Tun Anh, Trn Gia Hng, Nguyn Thu Yn, Trn Vn Tin (1998),
Mt s nhn xt v tnh hnh bnh si khu VTC min Bc 1996-1997, Tp ch
Y

hc d phng, 4 (38), tr 20-25.

10. http://vietbao.vn/suc-khoe/phong-va-dieu-tri-benh-soi/10890513/248/
11. http://vnexpress.net/GL/suc-khoe/
12. http://dichtehoc.com.vn
13. http://www.sinhhocvietnam.com/forum/showthread.php?p=8790

40

TING ANH
14. David A., Fuccilo, Jonh L. (1989), Measles virut, Diagnostic protein
incorporated into cell membrane, Intervirology, Switzeland, 11, pp.275- 281.
15. Norrby E., Gollmar Y. (1972), Infec. Immun., 6, pp.246.
16. Researchers Associates (1996), The National Institute of Health, Vaccine
handbook, Tokyo
17. Stanley A. Plotkin, Walter A . Orenstein (1999), Vaccines, 3rd edition, America.
18. WHO (1990), Guidelines fo r the preparation, characterization, and
establishment o f International and other standards and reference reagents fo r
biologycal substances, TRS no 800, annex 4.
19. WHO (1997), Manual o f laboratory methods fo r testing o f vaccines used in the
WHO Expanded programme on Immunization, WHO/VSQ/1997.04.
20. Norrby E. (1985), Measles, Virology (B.N.Fieds ed.), Raven press, New York,
55, pp.1305-1321.

41