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SEMINAR

A. T VN

Ngay t khi xut hin s sng, trong s u tranh ca cc loi sinh vt th bnh tt xut hin. c bit l s k sinh ca cc loi vi sinh vt ti cc ng vt bc cao gy ra nhng bnh tt him ngho to ra nhng nn dch thm khc v cp i nhiu sinh mng. ginh git s sng con ngi tm mi bin php nhm hn ch tc ng c hi ca cc i tng gy bnh. Vacxin c coi l mt tin b y hc quan trng nht ca th k XX Ngy nay cng vi s pht trin khng ngng ca cng ngh sinh hc con ngi tm ra c v kh hu hiu bo v chnh mnh vi s lng nhiu, an ton v nhanh chng. hiu r hn vn ny, chng em tin hnh tm hiu ti:

ng dng cng ngh sinh hc trong sn xut vacxin

Vaccine l g ?

Hng pht trin ca cc vaccine mi ?

Vaccine c tc
dng nh th

SN XUT VACCINE
Cng ngh sinh

no ?

hc ng dng
g trong sn xut vaccine ?

C s ca vic sn xut vaccine?

1.1. Vi nt v lch s v hng pht trin ca cng ngh vacxin

Th k 18, th gii chng kin ba i dch cm

Cc dch cm vn tip din trong th k 19, khi cc thnh ph ln dn ln v giao thng ng bin gip cho vic i li c thun tin - ng ngha vi kh nng ly lan tng hn. Dch cm nm 1837 nghim trng n mc Berlin, s ngi cht nhiu hn s ngi c sinh ra, v Barcelona th ton b cc hot ng kinh doanh b nh n.

Nm 1918 chng kin i dch cm nghim trng nht lch s th gii, thm ch cn c cho l i dch kinh hong nht trong cc loi bnh dch. Khong 50 triu ngi cht, trong ring Ty Ban Nha c 8 triu, v th dch cm ny mang tn cm Ty Ban Nha. Trong nh, bnh nhn cm nm la lit trong mt bnh vin Kansas,

Nm 1957 nh du dch cm chu . Nh cc tin b khoa hc, dch bnh nhanh chng c xc nh, cc bin php y t cn thit c trin khai, trong c vic s dng vc xin. Tuy vy s lng ngi cht v cm vn mc 2 triu. Trong nh, cc bnh nhn cm an Mch nm trong khu ngh tm nh thi u ca hi qun Copenhagen

Nm 2003, Bng n dch cm gia cm

Vic to ra nhng loi vaccine phng v cha bnh l thc s cn thit khi m dch bnh pht trin ngy mt phc tp v xut hin thm nhiu bnh nguy him

1.1. Lch s vacxin


Edward Jenner c cng nhn l ngi u tin dng vc-xin nga bnh cho con ngi ngay t khi ngi ta cn cha bit bn cht ca cc tc nhn gy bnh.
Nm 1796 ng thc hin thnh cng th nghim vc-xin nga cn bnh u ma, nhng nng dn vt sa b c th b ly bnh u b, nhng sau khi khi bnh, h tr nn min nhim i vi bnh u ma Edward Jenner

Louis Pasteur vi cc cng trnh nghin cu v vi sinh hc v min dch hc m ng cho nhng kin thc hin i v vc-xin.
Louis Pasteur

1.2 Khi nim v vaccine

Vc-xin l ch phm c tnh khng nguyn dng to min dch c hiu ch ng, nhm tng sc khng ca c th i vi mt s tc nhn gy bnh c th.

1.3. Nguyn l s dng vacxin


S

dng vacxin l a vo c th khng nguyn c ngun gc t vi sinh vt gy bnh hoc vi sinh vt c cu trc khng nguyn ging vi sinh vt gy bnh, c bo ch m bo an ton cn thit, lm cho c th t to ra tnh trng min dch chng li tc nhn gy bnh. Ni mt cch khc: s dng vacxin l to min dch ch ng nhn to

1.4 c tnh c bn ca mt vaccine

1.4.1 An ton
trng : khng c nhim cc vi sinh vt khc Thun khit : khng c ln cc thnh phn khng nguyn khc Khng c Liu s dng phi thp hn rt nhiu so vi liu gy c Tuy nhin cng c th gy ra phn ng ph mt s ngi
V
Phn

ng ti ch: Ni tim vaccine c th hi au, mn , hi sng hoc ni cc nh Phn ng ton thn: St thng, co git vi t l nh c th sc phn v vi kh nng xy ra him

1.4.1 Hiu lc
Vacxin c hiu lc ln l vacxin gy c min dch mc cao v tn ti trong mt thi gian di. Hiu lc gy min dch ca vacxin trc ht c nh gi trn ng vt th nghim, sau trn thc a. Trn ng vt th nghim

Cch 1:nh gi mc p ng min dch thng qua vic xc nh hiu gi khng th hoc xc nh mc dng tnh ca phn ng da. Cch nh gi ny cha cho bit hiu lc bo v, mi ch cho bit mc p ng min dch ca c th ng vt i vi loi vacxin th nghim. Cch 2: Xc nh t l ng vt c tim chng sng st sau khi th thch bng vi sinh vt gy bnh. Vacxin c tim chng cho mt cng ng, theo di thng k tt c cc phn ng ph v nh gi kh nng bo v khi ma dch ti.

Trn thc a (field test):

1.4 c tnh c bn ca mt vaccine


1.4.3 Tnh khng nguyn
L kh nng kch thch c th to thnh khng th. Khng nguyn mnh l khng nguyn khi a vo c th mt ln sinh ra nhiu khng th khng nguyn yu l nhng cht phi a vo nhiu hoc phi km theo mt t dc mi sinh c mt t khng th

1.4.4 Tnh min dch


Vacxin gy min dch bng mt vi khun hoc virus gim c lc, hoc vi mt protein c hiu c tnh khng nguyn gy ra mt p ng min dch To ra hiu qu khng cho c th v sau khi tc nhn gy bnh xm nhp vi y c tnh.

II. Phn loi vaccine


2.1 Cc phng php phn loi truyn thng 2.1.1 Vaccine ng lc (nhc c) hay vacxin sng

L vacxin ch t tc nhn gy bnh l gim tnh c bng cc k thut vt l, ha hc, hoc bng cc phng php khc nhau khng gy bnh nhng cn kh nng sinh sn. VD: Vacxin dch t vt ng kh

2.1.2 Vaccine bt hot (cht)


L loi vacxin m tc nhn gy bnh b lm cht bng phng php ha hc hay nhit . VD: VC XIN TU HUYT TRUNG HEO

2.1.3 Vaccine di n v hay vaccin thnh phn

L loi vacxin khng dng ton b t bo vi khun hay ton b virus m ch dng mt thnh phn c tnh khng nguyn ca chng.

3.1.4 Vaccine gii c t

L loi vacxin to ra do lm mt tnh c ca khng nguyn nhng vn c tnh khng nguyn.

2.1.5 Vaccine khng khng th idityp

L khng th to ra t cch dng khng th khng khng nguyn lm khng nguyn d lm vc xin cho tc nhn gy bnh

2.2 Vaccine tng hp


i.

ii.

iii.
iv. v.

Vaccine sng gim ng lc bng lc bng k thut di truyn Vaccine cha KN sn xut bng phng php ti t hp Vaccine vector Vaccine ADN Vacxin da trn c th chuyn gen. Sn xut cy chuyn gen nh Agrobacter tumefaciens

vi.

Vaccine bo t (spore vaccine)

Loi vacxin

u im

Nhc im C th gy bnh nh C th chuyn thnh dng gy bnh Virus c th b cht, cn bo qun lnh C th b nhim virus Cha genome virus c th gy bnh ung th Cn nui to lng virus ln, c th cn t cht b tr Tnh min dch c th khng cao C th phi tim nhc li nhiu ln Cha genome virus c th gy bnh ung th

n gin, d sn xut Vacxin Gi thnh r, tng i gim n nh ng Gy min dch ko di lc Khng cn tim nhiu (sng) ln n gin, d sn xut Gi thnh r, n nh Vacxin bt Khng cn bo qun hot lnh (cht) Kh nng nhim virus sng rt thp

Vacxin tng ph n tinh sch

Gy p ng min dch nh hng vi tng phn ca virus Khng tr li thnh virus ly nhim n nh, c th khng cn bo qun lnh

Sn xut phc tp, cn cht b tr Gi thnh thng cao Cn h thng nui thu lng virus ln Tnh min dch c th khng cao C th phi tim nhc li nhiu ln C th cha c ht virus hoc genome virus

Gy p ng min dch nh hng vi tng phn ca virus Vacxin Khng tr li thnh virus tch ly nhim dn Kh nng nhim virus sng g thp n nh, c th khng cn bo qun lnh

Nghin cu phc tp, chi ph cao Cn cht b tr Tnh min dch c th khng cao C th cn tim nhc li nhiu ln

Vacxin

Tc nhn gy bnh Virus vim gan B

Bnh

Loi vacxin

Do CDC gii thiu Vim gan B Vim gan Di n v bt hot

Bch hu/ho g/un vn

c t bch hu c t vi khun than Bordetella pertussis

Bch hu Than Ho g

Gii c t Gii c t Di n v bt hot Cng c th dng c t bo lm khng nguyn Di n v bt hot


Bt hot (Salk) Gim c lc (Sabin)

Haemophilus influenzae typ B


Poilo bi lit

Haemophilus influenzae
Virus Polio

Vim mng no Vim phi


Bi lit

Si/quai b/ rubella (MMR)


Thy u

Virus si Virus quai b Virus rubella

Si Quai b Si c

Gim c lc Gim c lc Gim c lc


Gim c lc Di n v bt hot Bt hot (c ht) Di n v bt hot

Virus thy u Thy u Vim phi

Vim phi Strepcoccus Pnuemococcus Pnuemonia Vim gan A

Virus vim gan Vim gan A (HAV) Virus cm Cm

Cm

Vacxin

Tc nhn gy bnh

Bnh

Loi vacxxin

III. C ng ngh sn xut Vacxin.


3.1 Sn xut vaccine truyn thng
To

sinh khi Lm bt hot i vi vaccine cc vi khun cht: C th d dng cc tc nhn dit khun (formalin, alcool, aceton, tia cc tm, siu m) i vi vaccine t vi khun sng gim ng lc Sn xut ra ch phm Kim tra sn phm

Sn xut vacxin tng hp


* C hai loi vacxin ti t hp gen: +> Vacxin ti t hp phn t: vacxin ti t hp phn t c to ra bng cch ghp gen hay np gen khng nguyn ca i tng gy bnh vo virus hoc vi khun. Virus hay vi khun ny sinh sn nhanh to ra nhiu phn t khng nguyn. Cc khng nguyn ny c tch chit, tinh sch lm vacxin ti t hp chng li cc bnh st xut huyt, vacxin chng vim gan B (HBV), vacxin chng bnh st rt, vacxin chng HIV

Sn xut vacxin tng hp

+> Vacxin ti t hp gen virus sng : Vacxin ti t hp gen virus sng l cc virus ng vt bin np cc gen biu hin protein thch hp mang tnh min dch t cc c th bnh l khc nhau. Vacxin ny c nhiu u im l : cng mt lc c th p ng min dch khc nhau trong cng mt c th, ng thi n c th ngn cn s khng thuc ca vi khun v k sinh trng gy bnh. Cu trc thng thng ca mt virus ti t hp bao gm: Mt l gen ngoi lai m ho protein khng nguyn ca tc nhn gy bnh Hai l virus ADN Ba l gen ch th

Sn xut vacxin tng hp

Quy trnh tin hnh ti t hp nh sau: Bc 1: Tin hnh xen on th cp ca b gen virus vo mt plasmid vi khun c mang gen khng li khng sinh (1) Bc 2: To t hp gm ADN khm c cha on khi ng (promoter) mnh gn vi gen khng nguyn v ui poliA. Sau xen vo vng ct hn ch ca virus.(2) Bc 3: l a (bin np) ng thi c plasmide v virus vo t bo ch nhn chun. Ti y virus ADN v plasmide hp nht v to ra virus ti t hp Bng phng php to vacxin ti t hp virus sng, ngi ta to ra vacxin u ma ti t hp cho ngi v vt nui, to vacxin adenovirus ti t hp chng mt s bnh h hp dng vin nhng ung.

3.2 Sn xut vaccine ti t hp


Mt on gen cn thit t phn t ADN ny

Mt phn t ADN khc (gi l th truyn)

ADN ti t hp

ADN ti t hp c a vo t bo vi khun c c tnh pht trin nhanh (TB ch)

T bo ch nhn ln

on gen cn thit cng c nhn ln

Phn t ADN ti t hp nhn ln nhanh chng

Bng cc k thut chit tch, tinh ch, ngi ta thu ly nhng phn t ADN mang on gen cn thit ny s dng vo nhng mc ch nh nh sn xut vacxin, thuc khng sinh

ng dng cng ngh gen trong sn xut vacxin


Hin nay c mt s vacxin c sn xut bng cng ngh ti t hp gen

Vac xin th h mi hay vacxin ti t hp gen

Sn xut vacxin l mm long mng th h mi


Tm on gen c tnh khng nguyn cao trong t bo virus Dng enzim sinh hc ct ly on gen ny

Chit, tch S dng ch to vacxin

Nhn ln bng cng ngh ti t hp gen

IV. Minh ha mt vi qui trnh sn xut vacxin 4.1. Vacxin c in c sn xut (VD: Vacxin vi khun thng hn)

Bc 1:Nhn ging: Nui vi khun thng hn trn mi trng thch trong 18 h sau dng NaCl ng trng ly vi khun v iu chnh t m c 5.108 vi khun/1ml. Cy vi khun ny vo mi trng mi sao cho th tch ban u vi khun chim 5% th tch mi trng Bc 2:Nui cy Mi trng lng casein cha 300-400 mg N ton phn, 200- 250mg% N amin, 0,1-0,6% pepton. Cy chm trong mi trng nui cy lin tc c h thng khuy vn tc 180-300v/ pht tng oxy cho h thng, h thng lc kh v trng vi lu lng 1lit/1pht. Nui trong 10-12 gi t c nng khong 6.1010 vi khun tng kh nng sinh sn ca vi khun thm dung dch glucoza 40%, gi pH= 7,6-7,8 trong sut thi gian nui cy .

IV. Minh ha mt vi qui trnh sn xut vacxin 4.1. Vacxin c in c sn xut (VD: Vacxin vi khun thng hn)

Bc 3: Lm bt hot Vacxin c lm bt hot bng cn: Ly sinh khi trn thn trng trong bnh c bit vi cn ethyl theo t l ln1:1:4, ln 2 1:10 Bc 4:Kim tra tinh khit v p ng min dch Kim tra tinh khit v p ng min dch ri a dung dch v im ng in bng NaCl c cha 0,25% phenol s to ta. 1ml vacxin cn cha 5.109 vi khun. vacxin vo ng v ng kh 400C- - 500C ri ng gi thnh phm. Vacxin ny c th nhp chung vi VI antigen tng hiu qu.

4.2. Quy trnh sn xut vacxin ti t hp(VD: vacxin cm)


Bc 1: Thu nhn mu bnh v thng tin tnh hnh dch t bnh cm Bc 2: Chn on, phn lp virus v cc phn tch s b Bc 3: Sn xut khng huyt thanh chn sng Bc 4: Phn tch c im di truyn v khng nguyn Bc 5: nh gi v chn la virus ng c lm vaccine Bc 6: S ti t hp virus cm bng k thut di truyn ngc

4.2. Quy trnh sn xut vacxin ti t hp(VD: vacxin cm)

K thut ny cho php 6 gene (m ha cho cc protein NP, PA, PB1, PB2, M, NS) ca virus cm A/Puerto Rico/8/34 (thng gi l PR8) kt hp vi 2 gene m ha cho protein HA v NA (ca chng khuyn co lm vaccine) c lm gim c lc to chng virus mi mang c im khng nguyn HA v NA ging vi chng khuyn co lm vaccine v c th pht trin trong trng nhanh kp thi cung cp vaccine khi c dch xy ra.

Minh ha k thut di truyn ngc (c in)

Minh ha vectors dng trong k thut di truyn ngc ci tin

Bc 7: Xc nh c im khng nguyn v di truyn ca chng ti t hp Bc 8: nh gi c tnh pht trin ca virus ti t hp Bc 9: Chun b cc ho cht cho vaccine bt hot

ngha vic sn xut bng cng ngh ti t hp gen


Nng cao nng sut sn xut vacxin To ra sn phm vacxin vi nhiu u im:
Vacxin ti t hp gen rt an ton v trong vacxin khng c s tn ti ca mm bnh Khng cn bo qun lnh nn gim c chi ph => ph hp vi iu kin s dng nhng nc ang pht trin

C.KT LUN

S ra i ca vacxin c ngha v cng to ln i vi nhn loi. N gip cho loi ngi thot khi nhiu i dch him ngho (u ma, bi lit; si, un vn v.v. Tuy nhin, nhiu bnh vn cn ang thch thc con ngi, cha c vc-xin no hiu qu ngn nga. Trong phi k nhiu bnh do k sinh trng (th d st rt, giun, sn), vi khun (lao), virus (cm, st xut huyt, AIDS v.v.). Mt s l do c th l cc tc nhn gy bnh bin i thng xuyn khin cho min dch khng cn hu hiu hoc thm ch tn cng ngay vo h min dch nh trng hp ca HIV v.v Cc loi vc-xin truyn thng ang dn mt i tnh nng phng bnh, c bit cn gy ra nhng phn ng ph cht ngi trong thi gian gn y. Tnh u vit ca cc loi vacxin ngy cng c khng nh: rt tinh khit, t phn ng ph, hiu lc to min dch cao, sn xut nhanh v gi thnh thp C th ni, nhng thnh tu t khoa hc k thut c bit l cng ngh sinh hc s m ra nhiu hng nghin cu mi v hiu qu trong cng tc iu tr bnh

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