TỶ LỆ MANG KHÁNG NGUYÊN BỀ MẶT VI RÚT VIÊM GAN B HBSAg TRÊN CÁC ĐỐI TƯỢNG KHÁC NHAU TẠI TỈNH LÂM ĐỒNG

Vinit Viem gan B HBV Iii lac nhan gay benh Viem gan B ",t nguy hiem.Nglloi mang Khang nguyen be m?1 virut Viem gan B Iii mol lrang nhungnguyen nhan lam lay Iruyen HBV, gay benh Viem gan

BỘ GIÁO DỤC VÀ ĐÀO TẠO

TRƯỜNG ĐẠI HỌC ĐÀ LẠT

HOÀNG CÔNG LONG

TỶ LỆ MANG KHÁNG NGUYÊN BỀ MẶT

VIRÚT VIÊM GAN B (HBSAg)

TRÊN NHỮNG ĐỐI TƯỢNG KHÁC NHAU

TẠI TỈNH LÂM ĐỒNG

LUẬN VĂN THẠC SĨ

NGÀNH: SINH HỌC

ĐÀ LẠT - 1995

80 GlADDuevA flAo T.o.O

m

HoANG CONG LONG

.£uiuL WL vI w.e cSi

VIRUT VIEM GAN B (HB sAg)

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CHUYEN NGANH: VI SINH vAT Y HOC

1"T'l·lr\I~·JI , ) , AI''''''I,"U,.

c6VAN KHOA HOC: GSTS HoANG THUY NGUYEN

PTS NGUYEN THU VAN

([)alal, 1995

El§ holm thanh Luiln an nay, toi xin chan thanh cam dn :

sinh Djch Ie hoc Trung Vdng - Ha NQi

• P.T.S NGUYEN THU VAN - Trung tam Ki§m dinh QuocGia - Ha NQi

che pham sinh hoc

• SS PHAN THANH HAl, Giam doc Trung Tam Chandoan Y Khoa - TP HCM

• TS LE sA DONG TrvClng Khoa Elao taO sau Elai hoc TrVClng Elai hoc DALAT

-• SS El6 HOU CHiNH - Giam doc Trung Tam Y hoc Dl,!phbng - Tlnh Lam Dong

- TTCElYK - TP/ HCM

Dich I§ Tay Nguyen

• TRUdNG ElAI HOC ElA LAT

Ela het long giup do va dQng vien

Virut viem gan BCut off value: gia tri ngtlang.

Negative control MeanGia tri trung binh cac chung Am

Positive: DtlClng tinhNegative: Am tinhAlpha FetoproteinHuman Immuno Deficiency Virus

Reverse Passive HaemagglutinationNgtlng ket hong cau thv d¢ng ngtl<;1c

Radio Immuno AssayMien dich ph6ngXI;!.

Enzym linked immuno Sorbent Assay

Mien dich Men

Counter immuno Electro Phoresis

Elien di mien dich doi Itlu

Primary Liver Cancer

Ung thtl gan Nguyen phatHepatocellular carcinomaUng thtl bieu ma te bao ganNational Institude of Hygiene ADN Epidemiology.Vien V e sinh Dich Ie hoc Trung UCing

World health organization

Hichuc y te the gioi

q.Jhdn LIlli Ilht1l

DAT VAN DE

Vinit Viem gan B (HBV) Iii lac nhan gay benh Viem gan B ",t nguy hiem.Nglloi mang Khang nguyen be m?1 virut Viem gan B Iii mol lrang nhungnguyen nhan lam lay Iruyen HBV, gay benh Viem gan sieu vi cap hoacman, sau d6 c6 the chuyen Ihilnh Xo gan va cuoi cung la Ung Ihll gan(carcinoma).

Hien nay Viem gan virut Iii mol van de dang dllc;Jc quan lam ti;li nhieu nllocIren the giOi Ty Ie nglloi mang virut nay aBac My va Chau Au Iliong doithap: 0,1 den 0,2% khong Vll(jl qua2% Song li;li nhieu nlloc ChauA, ChauPhi thi Iy Ie nay ral cao :15 den20% Ve phllong dien dich Ie hoc clla HBVral giong voi HIV

Thea 16 chuc y Ie the gioi (WHO) moi nam c6 kholmg 2 trieu nglloi trenloan cau chel do virul Viem gan B, hon 285 trieu ngllai mang mam benhtrang d6 nhieu nhat la ngllai ChauA -chiem Iy Ie75%

Ung Ihll gan va Xo gan la nhung benh hiem nghE!O gay tU vong nhanhch6ng sau khi c6 bieu hien lam sang, song neu phal hien sam agiai doandau, kel qua dieu Iri S8 101 hem rat nhieu £lay la nhung can benh tien Iriench"m, nhllng chac chan trong nhieu nam sau S8 c6 khong il ngllai bj Xogan dan cten Ung Ihll gan

Thang 3.1993, lrang Hoi IhllO Khoa hoc ve chuyen de Ph6ng chong Ungthll gan nguye~ phal ti;li Ha Noi, mot trong nhung muc tieu de ra la: lam Ihenao de pilat hien th"t sam Irong can bo, nhan dan nhung ngllai c6 cac yeu

to nguy co cao

Hien nay, Viel Nam dang n1lm trong vung dich tEl virul Viem gan B Iliu hanh

amuc do cao Lam £long la mol Tlnh mien nui phia nam Tay nguyen, c6Thanh pho 8a Li;lt la noi nghl mal Iy tllang cho du khach Neu chung lakhong c6 bien phap ng{m ch?n virul HBV S8 bien noi nay thanh 1 vungnhiem virut amuc do ngay cang cao £lieu nay S8 anh hllang ral Ion venhieu m?l: i<inh Ie, Xa hoi, Chinh tri, va £lai song clla ngllai dan song adayn6i rieng va clla ca nllOc n6i chung

Vi vf:!y, de tai se tien hanh dieu tra Huyet thanh va giam sat tinh hinh mangHBsAg l<;li Tlnh Lam EJong Viee ThlJ nghiem mau xae dinh virut HBV trenmoi ngLloi binh thLlong va tn§n nhung doi lLlong c6 nguy co eao bi nhiemHBV la dieu rat can thiel va d6 ehinh la mue lieu ella de ta;,

Xae dinh dLloe ty Ie cae nh6m ngLloi cae nh6m doi tLlong khae nhau e6

mang HBsAg DLlong tinh de sau d6 ve mf:!t Dieh te hoc se e6 nhung bien

phap h<;ln ehe eOng nhLl ngan chan st,llan truyen ella virut HBV trong daneLlt<;li cae Tlnh va Thanh pho trong ea nLlOe

Ngoai ra mue lieu ella de tai la ung dung cae tien b(> khoa hoc ky thuattrong chan doan de phat hien ngLloi e6 mang virut va dt,l ph6ng nhiem HBVdoi vol ngLloi am tinh

rph.iiJL title ltai

TONG QUAN TAl L1I;U

Viem gan B la mot benh do Hepatitis B virus (HBV) ral phb bien va t",athanh mot van de nghiem trong ve y Ie cong cong <'J nhieu nLioc tren thegioi ngay nay NhLlng benh nay con la mol moi de doa nghiem trong han:VinH gay ra Vi em gan B chinh la tae nhan gay Ung thLi thu ba dLlc;Je biet sauthuoe la.

BLioe di dau tien Irang viee xae dinh da dlic;Je thl/e hien vaa nam 1983 b<'JiBaruehs Blumberg t",i Vien nghien eLlu Ung thLi <'J Philadehphia trang khinghien cuu mot so Protein trang Huyet thanh ella mot benh ph€lm lay W

mot benh nhan bi benh L1a chay mau, ong da lam bi~u hien ro mot Khang

th~ phan ung voi mot Khang nguyen e6 trang mau mot thb dan Australia biViem gan

EJen nam 1968, Blumberg xae dinh d6 Iii Khang nguyen be m.;;t ella virutViem gan B

1 Virut Viem gan B:

Thuoc hO Hepadnaviridae NgLioi ta da xac dinh dLlc;Jc virut Viem gan B c6 5dau an mien dich :

- Khang nguyen be m';;t virut Viem gan B (HBsAg)

- Khang lh~ chong Khang nguyen be m';;t virul Viem gan B (Anti-HBs)

- Khang th~chong Khang nguyen 16i virut Viem gan B (Anti - HBc)

- Khang nguyen e Virut Viem gan B (HBeAg)

- Khangth~chong Khang nguyen e virut Viem gan B (Anti - HBe)

Trang 5 dau an mien dieh Iren ngLioi la thay dau an Khang nguyen be matvirul Viem gan B Iii quan Irong nhat (Hinh a)

HBsAg e6th~ton t",i trang mau bat eu lue nila <'J giai da",n nhiem cap, man,vii c6 th~ ton I",i suot dOi Nang do clla HBsAg DLiong tinh (+) giam danthea lubi nhLlng tae h",i ella Xo gan vii ung thu gan thi tang dan thea tubi <'Jnglioi e6 mang HBsAg (+) trang mau, thong thLiong Ung thLi gan chi xuathien sau mot thai gian tiem tilng W30 den 50 nam, nhLlng cOng nh';;n thaymot so ca carcinoma te baa gan <'J Ire em

CAU TRUC VIRUT VIEM GAN B

6 M(>I Nucleocapsid mang nhungkh ang nguyend~c bi$t

7 Enzym ADN - Polymeraza

8 B(> gen ADN clla HBV

HBV c6 mot eElU true hai Ian va protein dong tam Va boe,nghTa la va ngoai baa gam 3 protein dlioe gOi la "Ion hem",

"trung blnh" va "Ion" HBsAg, Khang nguyen be mat, e6 mattren ea 3 protein d6 Capxit hay va trong eau t",o blil motproteinIitrong vua baa boe vua tLidng tae voi ADN eua virut.

Bo gen eua HBV Ian dau lien dLioe ph an lap nam 1974 blii WilliamS.Roblnson IiTrLiong Y thuoe 8",i Hoc Tong HOp Stanford, Iii mot philn tU

ADN vong baa gam khoang 3200 Nuelectit 86 la bo gen nha nhat virutdong vat dLioe biet; bo gen (jenom) eua HBV eau t",o blii hai soi nhLing n6I",i e6 1 dae tinh bat thLiong : Ha; sol e6 ehieu da; khong biing nhau Chieudal eua sOi ngiin con gOi la sOi "dLidng" thi thay doi va chi biing 50 den 80

ehieu dai eua soi "am" Cau true dae biet nay la ket qua eua cd ehe nhanlen doc nhat dLioe vi rut su dung

Cau true vong eua virut dLioe baa dam blii SLi gMp do! giua cae baz Iim(Jtdau eua cae soi Vlrut HBV e6 eau true dae biet xit chat N6 baa gam e6 4gen dLioe gOi la gen S, gen C, gen P va gen X Chum phu len nhau rat

ehu ky nhan fen eOng baa gam trong cae do",n ghi rna

Gen S ghi rna hay chua dl,lng thong tin eho protein Ion hdn eOng nhLi ehoHBsAg Mot doan khoang 500 Nueleotlt Ii phia trLioe g"n S eOng e6 thedLioe phien rna eung lue (Wolfram H Gerlich) ta e6 the ph an blat trongdo",n nay 2 vDng Ii phia trLioe, tien S, va tien S2 tham gia vao Sl,l tong hopcae protein kMe eua va boe : Protein trung binh dLioe ghi rna blil tien S2 vagen S, Protein Ion dLioe ghl rna blii tien S" tien S2 va gen S VDng lien S,e6 val tro quan trong de vi rut xam nhap vao te baa gan (A Robert Neurath.eua Trung tam mau IiNew York da chung minh) (40)

Gen C ghi rna eho protein eua Capxit, eOng nhLi gen S, gen C eOng e6 1

do",n tien C ngiin IitrLioe gh; rna eho 1 pept;t ky nLioe tham gla vao viee liipghep h",t virut Gen P e6 ehieu dai tOi mue n6 eiit cae phan eua tat ea caegen, ghi rna eho cae Enzym eeJ ban eua ehu ky nhan len eua HBV Vi trieua gen X niim tran cae dau nol eua cae soi ADN eua Virut San phamProtein eua n6 kieh thieh sl,l the hlen eua tat ea cae gen khae biing tLieJng

Cac Hepadnavirul nhan len Iheo mot ceJ che bal Ihuang Ela phan vinHchua AON lai lao Iruc liep b(> gen cGa chung nho nhung Enzym gol laPolymeraza, cac polymeraza xli dung cac sol AON mol sinh lam khuonmau Iflp ghep cac soi mol bo sung Tuy nhien cac hepadnavirul sli dung 1phueJng phap glan tlep c6 st,l Iham gla cGa ARN lam Irung gian ceJ che nay

da duoc mo la Ian dau (;en nam 1982 blii Jesse W Summers va Williams.Mason cGa Trung lam Ung Ihu Fox Chase (Philadelphie)

CeJ che nay nhu sau: Khi m(>1 vlrut cGa hepadnavirut xam nhaP vao te baon6 dl loi nhan, Ii do nhung polymeraza te bao phlen rna Ihanh m(>t phantti

ARN da; Philn Iii ARN nay co 3500 Nucleolil dU(jc goi la lien jel10m TienJenom duoc boc b~ng m(>t AON polymeraza cGa virul lrong m61 capxit moi1"0 Ihanh vasau do Ira I"i nguyen sinh chal Ie bao 0 day, polymeraza cGavlrut lam cong viec phlen rna nguoc cGa tien jenom Ihanh m6t sol AON ammoi Elen khl soi am da hlnh Ihanh, tien jenom bj pM hGy bai cac Enzym,luc do cac AON - Pclymeraza co Ihe xay dt,lng s(jl dueJng sli dung soi amlam khuon mau Capxil va AON cGa virul cuoi cung duoc dal Irong mol va

boc ngoai moi khi virus roi Ie bao gan Khl vlrut duoc hinh thanh sol dueJngngung dai ra laP ltJc Elieu nay giai Ihich t<;ll sao chieu dai cGa s(jl dueJngthay do;'

Tal ca cac virul deu la nhung vi sinh val ky sinh Ie bao, cau 1<;10 blJl 1 Ihe

nhlem sflc la ARN hay AON (b6 gen) chua dt,lng trong 1 va boc protein nhu

da mo ta Ii phan tren Muon nhiln len vlrul phal dl vao Irong Ie bilO va xlidung b6 may cGa Ie bao de tong hOp cac protein cGavaboC va b6 gen cGavirul klli b6 gen da dU(jc bao Irongva boC thl cac hal virul moi roi Ie baova

c6 Ihe gay nhiem cac Ie bao Ian Can

CeJ che hap dan cGa st,l nhiln len cGa hepanavirul lam oM l<;Ii CeJ che slidung bal cac Relrovirul, nhu virul SIOA co b6 gen la ARN va sli dung m61philn tli AON lam Irung gian cac hepadnavirut con co nhung diem chungkMc nua vol Retrovirut ea 2 deu gay nhiem te bao m61 cach Ilem lang vakhong pha hGy Ie bilO do d6 Iht,lc Ie ta co the t11ay r6 hi en tuong nhlem virlll nhung khong xual hien Irieu chung, ceJ Ihe van hoan loan khae m<;lnh,sinh hO<;l1 van binh Ihuong, Iinh Ir<;lng nay co the keo daitU 8 - 10 nam Sau

do moi chuyen qua gial dO<;ln benh lam sang

Ta co Ihe Iham khao sl/ xam nhap va nhiln len cGa Relrovlrul duoi day(hlnI12)

3 PhOng ARN vil Enzym pllien

ma ngl1<;1c vila ben Irong Ie bilo

4 ARN cua virLiI

5 Enzym phien ma ngl1<;1c

6 ADN cua virLiI

7 Nilan Ie bilo Bach cau

8 ADN cua Ie bilo Bach cau

9 Te bilo san xuat ra nhieu ARN vaprolein virLiI

VinH HIV e6 eau 1<;10 don gilm, chI can 1 ph an tU ARN va mang Polypeplit.Chung ky sinh tren cae te baa B<;Ieh eau, te baa than kinh va 1 so te baakMe eua co thEi ngliai HIV xam nhap vao B<;Ieh eau bang each bam vilOmang B<;Ieh eau, ph6ng ARN vao ben Irong, can mang Polypeptid thi nam I!

ngoai Nha e6 Enzym phien ma nglioe ma ARN d6 tli nhan len thanh ADN,ADN nay gan voi ADN eua b<;leh eau trl! IMnh vat lieu di truyi'm moi, tU d6san xuat ra cae ARN eva HIV ma t<;lO tMnh 2 - 4 - 8 mam HIV kMe trongb<;leh cau Chung pM va vaeh B<;Ich eau roi I<;Ii bam vao cae b<;leh eau khaedEi tiep tue nay nl! va phat triEin Bang each d6 HIV pM hvy dan he thongmien dieh evacoIhEi ngliai

Thli tli va ehlie nang cae gen eva Retrovirut : gag, pol va env deu e6 thEi sosanh trlie tiep voi cae gen eva hepadnavirut : C, P va S (gag ghi ma ehoprotein eva capxit retrovirut, pol ghi ma eho Enzym phien ma va EnV ghi

ma eho cae prolein eva v6 boe) Ca 2 ho virut nay deu e6 anh hlil!ng doivoi ung thli

2 Sojlien quan giG'a HBV va PLC :

Cae nghien cliu Dich t8 hoe da ehling minh r6 rang la nhiem HBV man tinh

dv dEi gay Ung thli gan Cae dong vat c6 vu bi nhiE'im man tinh hepadnavirute6 Iy Ie mac Ung thli gan eao : tren 80% cae s6e maemot bi nhiE'im phattriEin mot Ung thli gan tien pMt (HCC) trang vong 2 nam Hans Popperthuoe El<;li hoc T6ng hop Cornell va John L Gerin thuoe El<;li hoc T6ng hOpGeorgelown da ehling minll rang e6 thEi gay ra Ung thli gan thlie nghiemtren s6e Maemot bang each tiem virut Viem gan rieng eva giong nay(30,40)

Cae Thl! nghiem nay da khi!lng dinh l<;Ii vai tro sinh Ung thu eva HBV va 10<;litru Sli can thiet cva cae dong yeu to eva sli phal sinh Ung thli C6 nhunghien tliong kMc e6 thEi tham gia vao benh I! ngliai: Thai ky tiem lang ditrlioc Sli pMt triEin eva Ung thli gan I! ngliai dai hall rat nhieu so voi s6emaemot va cae Ung thli thliang xual hien tren gan bixa. Cae chat gay Ungthlilrol1g moi trliang va slilieu thu qua nhieu rliou eung luon luon e6 thEineu len Irong sli pMt sinh Ung thli gan I!ngliai.(31)

Van decoehe gay Ung thli cva HBV van chlia dlioe giai quyet HBV e6 thEitrlie tiep gay ra sli phat triEin cae u Milt kMc, u eva gan e6 thEi xay denmot each gian tiep do Viem gan m<;ln tfnh, Xa gan va slitai t<;lO Ie baa xay

ra trong cae mo bi benh Nhieu virul ehiu traeh nhiem gay ra Ung thli mangcae oneogen, la nhung gen e6 thEi lam bien d6i trlie tiep cae Ie baa binhiE'im eva co thEi (Nhung oneogen tliang \lJ eung dlioe tim thay trong cae

Ie baa binh thliang trong d6 chung kiEim soal sli sinh trlil!ng va pMt triEin)tuy nhien bo gen eva HBV hinh nhli khong mang oncogen Them vao d6,

thai gian tiem tang dai giua lue nhiem HBV tOi lue xuat hi$n u khong phu

hOp voi ykien eha fang mot oncogen eua virut ch(u trach nhi$m gay Ung

thLi gan Tuy nhien, ngLiai ta e6 the quan niem riing Protein hoat h6a caegen san xuat bai gen X eOa HBV e6 Ihe can thiepVilO mot giai doan sam

eOa qua trlnh phat sinh u

onLiae ta PLG la Ung thJ dung himg thu 3 sau Ung thLi pheii va Ung thLi daday (pheii : 22.5%; Da day 15%; Gan 12%) Gae eong trinh nghien euu trenthe giai da chung minh moi lien quan chat ehe giua nhiem HBV va PLG.Nhiem HBV man linh dLioc xem la nguyen nhan quan trong nhat gay HGG.Elae biet (j Ghau A va Ghau Phi (S.Z.Yu T.LSung Ma Kew, T.G Tan1991) chi diem de tiep can nhat eOa nhiem HBV la sl,! e6 m;;;t eOa HBsAgtrong Huyet thanh.(7,12,13,26)

Mot so cae eong trinh eOng da chung minh: Huyet thanh ngLiai e6 mangHBsAg (+) nguy e(J tLiang doi de dang m;:'c HGG eao gap 223 Ian so vai

ngLiai Am linh (Beasley-1981)

3 Cae vung d!eh Ii! Iren Ihe giai

Thea tei ehue y 1e the giai (WHO) thi hien nay tren the giai e6 3 mue do

(1) Vung dieh tEi nhiem HBV thap: (0 cae nLiae Tay Au, B;:'e My): Ty Iemang HBsAg (+) VaG khoang 0,2 - 0,5%

(2) Vung dieh te nl1iem HBV trung gian: 0 cae nLiae Elong Nam Ghau Au,Trung Elong: Ty Ie mang HBsAg (+) tU2 - 7%

(3) Vung dieh te nhiem HBV eao: 0 cae nLiae nhLi Trung Quae, £long Nam

A, GhilU Phi Ty 1$ mang HBsAg (+)tU 8 - 20%

Ngoai ra con e6 mot so quae gia niim trong nhung vung dieh te nhiem HBVrat eao, ty Ie mang HBsAg (+) tren 20%

WHO eOng eho riing 3 10ai ty Ie dau eOng dO tap hOp dai da so cae nLiae

Nhung euoe dieu tra ve ty Ie nhiem HBsAg dLioe tien hanh trong khu vl,!e

cae dao(j Nam Thai Binh DLiang eho thay nhling bien deii nhieu khi rat Ian

ve ty Ie mang HBsAgtU quan dao nay sang quan dao kia ho;;;e tU dao naysang dao khae va ngay ea trong eung mot dao(3) Tham khao bang sauday ta se thay r6 nhung bien dong nay

Muc dO d[ch Viem gan B ILlu hill1h dia phLiong IJNam Thai Binh DLiong.

NLide - Dao hay quan dao

Societe, Tuamotu

HBsAg (+)

< den 7%

MuedQ virutlLiu hanhTrung gian

Niim1989

Tim Caledoni (Grand Terre)

Tan CalMoni (Grand Noumea)

198119871986

Tu van de tren ngLiai ta khuyen cao: Tat ca nhan vien b$nh vien ca nhungngLiaitNctiep hoac khong trvc tiep lam ve V deu c6 nguy co cao bj lay nhiem

vi rut Viem gan B Do d6 tat ca can phai dLic;JC tiem chUng (18,25,29)

Viet Nam nam trong vung c6 muc dO ILiu hilnh virut HBV cao, theo nhungbao cao bLicJc dau muc dO nhiem virut tU 15 den 20% tuy Iheo tUng d[aphLiong T<;ii Ha Noi theo tac gia Hoang Thuy Nguyen va Nguyen Thu Van(Vi$n V$ sinh Djcht~ h<;>c Ha Noi)(5,6,9):

- NgLiai cho mau c6 HBsAg (+)

- NgLiai khoe m<;inh c6 HBsAg (+)

- B$nh nhan Viem gan c6 HBsAg (+)

- B$nh nhiin khong phai Viem gan c6 HBsAg (+)

T<;ii thanh pho He, Chi Minh thea lac gia Tran Van Be (Trung tam truyenmau va huyet thanh):

- NgVai cho va hien mau c6 HBsAg (+) 9,5%

- Benh nhan Ung thV gan c6 HBsAg (+) 61,8%

Theo tac gia Ngo Quang Lvc (Ha Noi) Ty 1$ ngVai cho mau c6 HBsAg (+) la

9 - 10% Mot nghien cuu khac tren nh6m doi tVc;Jng la nhiin vien y te IJ HaNoi la: 9,83%

Va theo tac gia VO Tri$u An, VO Thuy Hien (16), thLJ nghi$m tren 43 nguoitrang d6 Nhan vien Y te, Nhan vien Philng xet nghi$m sinh hoa, B$nh vi$nNhi va m¢t so can b¢ thu¢c ph6ng mien djch b¢ men sinh Iy b$nh de c68/43 truong h<;lp HBsAg (+), chiem ty 1$ 18,6%.

Tren nh6m phu nv c6 thai ty Ie mang HBsAg (+):

• Vi$t Nam (Hai ph6ng)(11, 15) 12,59% (32 HBsAg (+) /254 ba me)T?i Portugal (23): Ty 1$ nguoi kh6e m?nh nhiem virut Viem gan B la 24,3%(1984)

Ta c6 the tham khao ban do Dich te hoc virut Viem gan B dudi day

Vung Djch te nhiem HBV thap

• Vung Djch te nhiem HBV trung gian

• Vung dich te nhiem HBV cao

4 Nguy eel nhiem HVB :

Tal ca nhung nguoi chua m<'lc b~nh Irong cong dong deu co Ihe nhiem vinH(24,32) Trang so do cac doi luc;1ng sau day co nguy cd cao hdn:

• Tre sinh ratU nguoi me co HBsAg Dudng Hnh Tren 70% cac Ire sd sinhnay se Ira Ihanh nguoi mang mam b~nh. Khi Ion len chling co nguy cd bj

Ung Ihu gan gap 200 Ian so voi Ire khong mang virli!

• Nhung nguoi Irong gia dinh Nhung nguoi liep xlic Ihan mal voi nguoi coHBsAg Dudng Hnh

• NMn vien y Ie, cac ky Ihual vien phong xel nghi~mva nhung nguoi lamcong lac nghien cuu thuong xuyen liep xlic voi mau va cac che philm tU

• Nguoi chich xi ke, ma lliy

• Nguoi xam minh, xo10lai hay b~nh nMn duc;1c dieu Iri biing cham cuu

• Khach du Ijch Ihuong xuyen den vung dich

5 51! lay Iruy;;n vii lien trien eua nguoi mang virlil Viem gan B

• 51! fay 'ruyen:

Eluong lay Iruyen clla vi rlit HBV la qua cac D[ch sinh Iy cua Cd the nhu:mau, nuoc bol, linh dich, d[ch am dGlO va qua cac vel rach clla da Gho nenst/ liep xlic chal cile hay giao hc;1p co Ihe Iruyen HBV Elieu nay gilli thichduc;1c st/ lay Ian virlil Vi em gan B cao lrang n¢i b¢ m¢1 gia dinh, hay m¢tc¢ng dong nho (34,35)

• ruin triifn ella nglf6i mang virtU Viem gan B.

NHIEM VI RUT VIEM GAN B

VIEM GAN CAP

VIEM CAN DQT PI-JAr

IrtJong hQpl

~ -1.Elia dii!m :

Cac mau mau dllc;Jc lay tV cac diem dEW cll Phan bo deu trong Tlnh Lam

£)ong, tVcac B$nh vi$n T]nh, cac Cc;JSO Yte trong Tinh va Thanh pho, cacBenh vi$n Huy$n v.v

2 Eloi tliong :

Gam cac Nh6m nglloi nhll sau :

a Nh6m kh6e m;;lnh: (can bo cong nhan vien, sinh vien, nhEw dan )

b Nh6m benh nhan Viem gan: (Sau khi d8 c6 chtin doan lam sang)

c Nh6m benh nhan khac: (Gam cac b$nh nhan den ph6ng kham cua cacBenh vien hoac dang dieu tri t",i cac khoa)

d Nh6m benh nhan Xc;J gan: (sau khi c6 chtln doan Lam sang la xd gan).

e Nh6m Ung thll gan: (Sau khi da dllc;Jc chtin doan Lam sang va xet

nghi$m Alpha - fetoprotein Dlldng tinh)

f Nh6m cho mau: (T",i B$nh vi$n Tlnh Lam £)ong va Benh vi$n II BllO Lac)

3 Lay mau vaxU'II' mau :

Lay mau tlnh m",ch 2ml, sau d6 tach chiet huyet thanh Trllong hop Ironghuyet thanh c6 hongdIU bi va phai lam s",ch bAng iy tam Tat ca cac mauhuyet thanh sau khi tach neu chlla dung ngay phai bao quan I",nh tV +2den +8°C Muon bao quan lau hdn phai lam dong I",nh (-20°C)

4 Ky thu~tphat hien Khang nguyen be m~tvirut Viem gan B

a Khao sal lam sang:

Khong thay vang da, vang mal tru b$nh nhan Viem gan , Xd gan va Ungthll gan

b Khao sat mau:

- Ky Ihu(it SERODIA - HBs

· Kit SERODIA - HBs clla Hang Fujirebio inc (Nh(it)

- Ky lhu(it ELISA (Enzyme linked Immuno Sorbent Assay)

· Kit micro Elisa HBsAg clla Labo Viem gan - Vi$n ve sinh Dich tehoc Ha Noi

· Kit 1M x HBsAg cua hang ABBOTT Laboratories (USA)

1 Ky lhu~lSERODIA - HBs :

SERODIA - HBs Ii Phliang phap Ngling ket hong cau thu dong Ngli<,1C(Reverse Passive Hemagglutination test) Dung de xac dinh Khang nguyen

be mat virut Viem gan - B

a. NguyenIy eua thii' nghiiim:

Bil Sinh ph am HBsAg - test gam cac thuoc thu cho Thu nghi$m nglingket hong cau thu dong ngli<,1c dung de xac dinh Kh8.ng nguyen be mi;ltvirut Viem gan - B (HBsAg) trong huyet thanh, huyet tliang nglioi, dl,latren II' thuyet Hong cau cliu ho",t h6a dli<,1C gan voi Globulin mien dichAnti-HBs (lgG) tho da dli<,1C hap phu c6 do tinh s",ch cao va ngling ketmot cach di;lc hieu khi c6 mi;lt HBsAg trong huyet thanh, huyet tliangnglioi

Thu nghiem nay c6 mot so liu diem sau :

- So voi cac Thu nghiem thuoC thehi) 1 va 2 nhli Khuyech tan tren gel.8i$n di mien dich doi lliu thi Thli nghiem nay c6 do nh",y cao han

- Quy trinh thl,lc hi$n don giim

- Tiet kiem thai gian 80C ket qua sau it nhat la 1 gio va doc biing matthliong

b. 89sinh phii'm:

Bil sinh ph am Serodia HBsAg bao gam cac thuoc thu:

- Control celis: Kh8.ng HBs - Hong cau cliu ho",t h6a - £long kho, Hoannguyen biing dung dich d$m trlioc khi dung

- Antibody sensitized cells (D",ng dong kho)

- HBsAg chLlng dLiang (Positive control) : chlia bat ho"t

- Phien nhl,la day hinh chCi "U"

- Que pha loang 25;t1 (#0,025 ml)

- 6ng nhl> gi<;>125~d1gi<;>t.

Khong dung cac ong nhl> giot khac

- Micropipets (0 -50~d);pipet 1cc

- Dung dich glutaraldehyde, hoilc sodium hypophosphate

d Thl/chi~nthynghi~m:

- Gi<;>t 2 gi<;>t (# 50~1 )dung djch d$m VilOtrong gieng Iva 1 gi<;>t (# 25~1)vao gieng II

- Dung Micropipet cho5~dhuyet thanh vao gieng I

- Dung que pha loang (Io"i 25~d) d~ tron deu, Huyet thanh trong gieng 1

se c6 dO pha loang la 1/10, cung dung que pha loang d6 chuy~n quagieng II (Ela c6 siln 25~1 dung dich d$m) Tron deu, Huyet thanh trong

gieng II luc nay c6 do pha loang la1/20.

- Nhl> mot gi<;>t (25J.l1) khang HBs hong cau cliu ho"t h6a (Tli d"ng dongkho dil hoi chlnh trtidc d6 30' va bao quan CJ +2oC den +1O°C) vaogieng I, va 25J.l1 Ie bao khang the nh"y cam vao gieng II Nhti vaY nang

do cuoi cung cua Huyet thanh sa la:

1/20 trong gieng I 1/40 trong gieng II

- Song song tien Mnh mau Chung Am va mau ChL1ng Dtiong, dung d$mmuoi phosphate lam ChL1ng Am

- Anti - HBs Hong cau cliu ho"t h6a (Control cells) (J.ll) 25

- Elo pha loang huyel thanh cuoi cung 1 20 1 : 40

e. Nh~n dinh kif!qua:

- Eloc kat qua sau it nhat la1gio, doc bang mat thllong

- Tuy vao muc do Ngllng kat rna ph;m bi$t : Am tinh va Dliong tinh

- Hoi;ic ngam vao dung djch Glutaraldehyd2% it nhat la1 gio

- Hoac dung djch Hypophosphate :1gio

- Hap lIdt (Autoclave) :121° C /gio

a. Nguyen fy cua Thu-nghi~m :

Thu nghiem cua HBsAg Micro-Elisa la mol Thu nghl$m tn,/c tiap, khongcl;lnh tranh de xac dinh Khang nguyen be mat vinH Viem gan - B, dungkhang the don dong khang HBs la khang the gan ban de phu cac 10 cuaphian nhlla va khang theW huyat lhanh de khang HBs lam cong hop vdlEnzym Peroxidaza

: HBsAg

Mau din Thli nghiem dlic;1c u voi Khang the giln ban trong 10 nhl,la Sau khirlia 10 nhl/a de 10<;1i b6 cac chat kh6ng giln, cho them c(>ng hop khang themen va luc do S8 xay ra phan ung giua c(>ng hc;1p khang the men nay voiphuc hc;1p Khang the· Khang nguyen da hinh thanh trong Ian u thu nhat,sau khi ph an ung ket thuc va nia Ian thu 2, tien hilnh u tiep voi dung dich

co bim cua men va chat hien mau, luc do mau S8 xuat hien neu mau Thlinghiem la Dliong tinh voi HBsAg

Neu kh6ng thay mau xuat hien co nghia la mau Thli nghiem Am tinh

b Cac thuo'c thu kern theo: (cho 96thunghi~m)

· Phien dEW dung cho chuan d(> vi Ilic;1ng

· M(>t phien dec (8 x 12 10) giln khang the don dong khang HBs.

· Chat bao quan: Merthiolate

· Dung d[ch dem co chat (2 x 3ml)

Dung dich dem Phosphate • citrat dac 10 Ian co chua Hydrogenperoxyde Khi dung pha loang 10 Ian

· Chat hien mau (8 x 5ml)

· 1,2 Phenylenediamine Dihydroclorid tinh the

· Chung dliong ( 1x 1,5ml)

· HBsAg nglioi pha loang trong huyet thanh nglioi thliong

· Chat bao quan: Natri azit

· Chung am (1 x 1,5ml)

· Huyet thanh nglioi thliong pha loang khong chua HBsAg

· Chat bao quan: Natri azit

· Dung d[ch dem cacbonat (2 x 6,5ml)

· Dung d[ch dem carbonat dung de hoan nguyen khang the giln ban

· Chat bao quan: Natri Azit

c. Cac nguyen v~tIi{iu can thilt:

H$ thong rua ban neu r11a bang tay chu ytranh khOng cham to; be matben trong cac 10nhl,ia.

Mot may doc ban c6 bl,idc s6ng 492nm

d Thl/Chi~n Thlrngh;~m:

Chuan bi cac thuoc thu •

1 Dung d!ch nla: Pha loang dem muoi Phosphate d,;,m dac (Thuoc thu3) UO vd; nl,idc cat Cho mot phien nhl,ia 500ml la duo Dung dich rua

da pha loang nay ben vung trong 2 tuan IJnhi$t dotU+2 den +Soc

2 Ci)ng h<;1p: Them 5ml nl,idc cat 2 Ian vao 10 (Thuoc thu 2), dE! dungd[ch 10' kh6ng lac va sau d6 lac nhe cho deu Dung dich cong hOp dapha ben vung trong 2 tuan IJnhi$t do +2 den +SoC

3 Chua'n b! dung d!ch hi$n rnau cd cha'1: Trong khoang 5 - 10' cuoicung cua Ian u thu 2, tlnh Il,iong dung dich mau Cd chat can thiet CUmol 10 chat hien mau (Thuoc thu 5) cho them 5 ml dung dich d$m cd

chat da pha loang 1 : 10 (Thuoc thu 4), bao quan tOi

4 Dung dich d$rn cdcha't: Pha loang dung dich d$m Cd chat d,;,m dac(Thu6c thu 4) vdi nudc cat (1 : 10)

5 Chung: So iuong chung am duong va chung dE! trong phV thuoc vao

so hang10 nhl,ia xu dVng cho mol Ian Thu nghi$m (xem bimg 1).Bang 1:

Tom tat qua trinh ph an ung nhtl sau :

a Cho dung dich khang the gan ban