V i e t n a m J o u r n a l of Physiology 2 1 ( 2 ) , 6 / 2 0 1 7 I S S N : 1 8 5 9 - 2 3 7 6
LfNG D g N G K V T H U A T ARMS - REALTIME - PCR D i XAC DjNH DA HINH GEN CYP2C19 VA ITGB3 LI^N QUAN B^ K H A N G CLOPIDOGREL VA ASPIRIN
& BENH N H A N MACH V A N H
N g u y i n Thj Minh N g o c \ Tri^u Tien S a n g ^ N g u y i n Thj Trang^
TOM TAT: Hien nay, b$nh m§ch vanh la nguyen nhan chinh gay td- vong vd tdn phe a cac nuKK phat triSn. Nhieu nghien cu-u da chi ra cac da hinh CYP2C19*2, CYP2C19''3 gen CYP2C19 va PIA1/PIA2 (Leu33Pro} gen ITGB3 co lien quan din su- de khdng clopidogrel vd aspirin. DNA du%fc tdch tCr 40 benh nhdn se duKvc su- dung ky thu$t ARMS- real - time PCR di xdc dfnh cdc da hinh gen CYP2C19*2, CYP2C19*3 vd PIA1/PIA2 (Leu33Pro). Khi phan tich da hlnh gen CYP2C19 (CYP2C19*2 vd CYP2C19''3) phdt hien 21 ngircd (52,5%) c6 mang dot biin gen, frong do co 14 ngwai (35%) Id mang gen dj hop tu' va 4 ngir&i (10%) mang ddng hap td" ddt biin CYP2C19*2 va 3 ngieai mang gen di hap CYP2C19*3. Vai da hinh PIA1/PIA2 gen ITGB3 phat hien 4 ngwal mang gen di hgp tir (10%). Nhw vay, ty 1$ phat hien cac da hinh gen tren kha cao a nhdm benh nhdn dieu tri kep clopidogrel vd aspirin. Trong bdi bao nay, chung toi hoan chinh quy trinh kT thuat ARIi/1S - Realtime PCR vd img dyng di xac dinh cdc da hinh gen CYP2C19 vd ITGB3 frong chin doan tinh khdng thudc clopidogrel vd aspirin ir bdnh nhan mach vanh
Tip khoa: BSnh mach vanh, CYP2C19, khdng clopidogrel, 1TGB3, khang aspirin.
I.DATVANDi sau chCcng minh 3\eu nay: Nghien cuu BSnh mach vanh (BMV) la nguydn CARRIE (1996); CURE (2001), PCI-CURE nhan chinh gay ttf vong vS tan phe d' cac
nud'c phat trien. Mac du, benh mach vanh tan suat till' vong cd giam trong 4 thgp ky trud'c tai My va cac quoc gia kh^c, nhung nd v i n chiem 1/3 nguydn nhan t u vong chung d' nhung ngud'i tren 35 tuoi [1], ud'c tfnh cd khoang mpt nOa nam gid'i va n u gid'i tuoi trung niSn tai My cd bieu hien BMV [2]. Tai Vi§t Nam, BMV dang ngSy cang gia tang ro ret gay nhi^u thay d6i trong md hinh benh tim mach.
Trong thyc hanh lam sang clopidogrel 1^
thudc chong ket t$p tieu cau lya chpn thCr hai sau aspirin, nhung hi#n nay cung cd nhi4u nghien CCFU cho thay v\&c k^t hgp dong \ha\ hai loai thuoc nay trong mpt so tinh huong chon Igc cd hieu qua cao han vi§c dung dan dgc [3]. Mpt so nghidn ci>u ' Ogi hgc Y HA N6i
^ Hpc vien QuSn y
* Tac gia chju trach nhi^m chinh:
NguySn Thj Trang
Email: [email protected] Ngay nhan bai: 15/5/2017 Ngay ph5n bi#n: 16/5/2017 Ngay chip nhSn dang 20/6/2017
(2001); CLARITY-TIMI 28; COMMIT-CCS 2.
C a che de khdng aspirin va clopidogrel hidn nay dup'c nghidn c u u va chap nhan nhieu nhat la da hinh kieu hinh gen di truyen. Nhieu kieu da hinh di tmy^n da dug'c xac dinh c6 lidn quan vd'i d ^ khang aspirin nhu da hinh gene C O X 1 , da hinh PLA1/A2 cua gen ma hda GPIIIa cua gen ITGB3, da hinh gen quy djnh thy t h i ADP P2Y1 tren tieu c l u , vWF va thy the collagen...[4]
V a i de khang clopidogrel, da dang vd kieu hinh cua gen CYP2C19 dug'c de cap nhieu nhat. Cd it n h i t 17 kieu alien cua gen nay lien quan vd'i k i l u hinh cd the lam tang, giam hoac cSn bang R-130964 lidn quan den typ hoang dai Allen cua typ hoang dai dug'c ky higu *1 va kieu gen hoang dai ky hidu CYP2C19*1/"1 hoat dpng binh thud'ng. CYP2C19*2 va CYP2C19*3 khdng cd chuc nang, cac kieu alien khac lien quan den bat hoat hoac giam c h u y l n hda thuoc nhu CYP2C19'4,*5,*6,*7 v& *8, tuy nhien cac
V i e t n a m J o u r n a l of Physiology 2 1 (2), 6 / 2 0 1 7 I S S N : 1 8 5 9 - 2 3 7 6
kieu alien nay t i n suat xuat hien it han CYP2C19 ' 2 va *3 Id k i l u gen chinh gdy giam chCpc nang alien d' ngud'i da trang (85%) va ngud'i chdu A (99%). Nhi>ng benh nhan cd tinh trang chuyen hda thu6c kem do m i t chuc nang ca 2 alien, tinh trang c h u y l n hda trung gian khi chT mat 1 alien. Benh nhan de khang aspirin hay de khang clopidogrel do da hinh di truyen can s u dung cac thu6c khang t i l u cdu khac trong phdng ngira nguyen phat va thu phat bdnh ly tim mach. K i l u gen n&y phy thudc n h i l u vao chung tpe [5]. Chinh vi vay vide phat hidn nhanh kieu gen lidn quan de khang aspirin va clopidogrel bang ky thuat PCR la rat can thiet.
Muc tidu nghien cO>u: Hoan chinh quy trinh va Crng dyng d l xac dinh cac da hinh gen CYP2C19*2 (G681A), CYP2C19*3 (G636A) va Leu33Pro (T1565C) gen ITGB3 bang ky thuat ARMS - PCR d- benh nhan mgch vanh da dat stent
2. D 6 | TU'O'NG V A PHU'ONG P H A P NGHIEN CLTU
2.1. Doi tiFp-ng nghidn cu>u: 40 benh nhdn mach vanh ( d u y c chan dodn mac hdi chiing vanh cap: nhoi mau ca tim, dau that nguc khdng 6n djnh) da d u g c dgt stent DMV qua da va co chf djnh d i l u trj clopidogrel va aspirin tai Khoa Khdm benh - B#nh vidn Bach Mai tir thang 6/2016- 12/2016.
Nghidn c u u xac djnh dot bien gen dup'c thyc hidn tgi Trung tam t u van Di t r u y l n - Bdnh vidn Dai hpc Y Ha Ndi.
2.2. Phu'ong phap nghien ci>u : DNA dup'c tach c h i l t t u mau ngoai vi (chong ddng EDTA) bang kit tach DNA Express cua hang Lytech (Nga).
S u dung ky thuat ARMS - Realtime - PCR de xdc djnh cac da hinh gen CYP2C19 (CYP2C19*2 va CYP2C19*3) va PIA1/PIA2 (Leu33Pro) gen ITGB3 v6i trinh t y mdi (Lytech, Nga):
Bang 2 . 1 . Trinh t y moi Trinh t y mdi CYP2C19*2
(G681A)
CCCACTATCATTGATTATTTCTCG - 3' 5 ' - CCCACTATCATTGATTATTTCTCA-3'
- ACGCAAGCAGTCACATAACTA - 3' CYP2C19*3
(G636A)
- GGATTGTAAGCACCCCCAGG - 3' 5 - GGATTGTAAGCACCCCCAGA - 3'
- TCACCCCATGGCTGTCTAGG - 3' Leu33Pro
(T1565C)
- GCTCCAATGTACGGGGTAAAC - 3' - GCTCCAATGTACGGGGTAAAT - 3'
RP 5' - GGGGACTGACTTGAGTGACCT - 3'
Ghi chu: FP - FonNard primer, ASP - Allele specific primer, RP- Reverse primer.
KT thuat ARMS - Realtime - PCR dug'c thyc hien tren may CFX96 (BioRad, My) vd'i chu trinh luan nhiet: 93°C - 1 phut, 35 chu ki (93°C - 10s, 64°C - 10s, 72°C - 20s), 72°C - 5 phut.
Sir dung bd kit DNA - express de tach c h i l t DNA va ky thudt ARMS - realtime - PCR nhan gen va phan tich dot b i l n la phuang phap t i l t kiem thai gian: trung
binh 45 phiit cho tdch chidt DNA va 90 phut cho xac djnh da hinh gen. N h u vdy kT thuat nay cd kha nang ung dung cao trong thyc hanh ldm sang de xac djnh nhanh vd chinh xac kieu gen cua benh nhan trudc khi dua ra phac dd cung nhu l i l u d i l u trj clopidogrel vd aspirin.
ADN cCia 10 benh nhdn lya chpn ng§u nhien dup'c phan tich dot bien bang ky
V i e t n a m J o u r n a l of Physiology 2 1 ( 2 ) , 6 / 2 0 1 7 I S S N : 1 8 5 9 - 2 3 7 6
thuat giai trinh t y gen Sanger.
Cac so lieu d u a c thu thap va x u li tren ph^n mem SPSS 16.0.
2.3. D g o du>c n g h i e n c i r u : Cac s6 lidu vd thong tin nghidn c u u ta hoan toan chinh xac, trung t h y c va khach quan da dyi?c xac nhan va chap thudn bd'i ca sd' nghidn ct>u. Nhung thdng tin ve bdnh nhan dup'c gii> bi mgt va chi phyc vy cdng tac nghidn ci>u.
3. K^T QUA
3.1. Hoan c h i n h q u y t r i n h k y t h u g t
A R M S - Realtime PCR
Toan bg tong s6 40 m i u nghien c u u dup'c tach ADN bang phuang phdp ADN- express (thyc hidn trdn mdy Lytech, Nga), tuy nhidn chung tdi giam the tich mau vd dung djch tdch ADN so vO\ protocol hud'ng dan cua hdng. Cy the, lug-ng mdu ngogi vi la 2 0 0 M I va dung djch tach ADN la 200iJl, thay vi 550pl mdu ngogi vi va 550 pi dung djch tach ADN nhu trong protocol.
T i l n hanh kiem tra do tinh sach dgt chuan thu dup'c ket qua sau
Bang 3 . 1 . K i t qua do nong dp va theo phuang phap ADN'
dp tinh sach ADN
•express Mau
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
N 6 n g d o A D N D o t i n h s ^ c h ng/pl
795,2 948,8 616,1 554,9 831,8 633,3 1006,3 1199,0 683,9 876,3 830,4 849,5 888,7 828,3 310,9 906,1 515,3 446,7 671,8 453,5 N o n g d $ DNA t r u n g 1
693,51 ± 227,94 1,9 1,75 1,94 1,84 1,70 1,87 1,91 1,82 1,79 1,73 1,75 1,83 1,86 1,81 1,69 1,88 1,80 1,72 1,76 1,89 binh
IVIau 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40
N o n g d o AND ng/Ml 1117,3 553,6 903,3 386,9 640,6 345,1 903,7 855,4 298,6 367,9 543,6 567,8 609,2 773,8 467,2 1016,8 804,5 678,6 588,9 470,7
D o t i n h s ^ c h 1,78 1,83 1,79 1,20 1,88 1,95 1,86 1,77 1,74 1,83 1,90 1,82 1,75 1,94 1,76 1,84 1,95 2,0 1,83 1,71 D p t i n h s ^ c h t r u n g b i n h
1,81 ± 0 , 1 2
Nhan xet: Lu'gng ADN thu du'O'C tu- cac m l u t u c n g doi cao va on dinii vd'i nong do cao nhat dat 1199,0 ng/pl, thap nh^t dat 298,6 ng/^jl; dp tinh sach cao nhat la 2,0 th^p nhat ia 1,70. Nhu- vay, vi$c giam t h i tfch i<it tach
va the tich mau so vo'i protocoi hang cung cap vira tiet l<ipm dtrpc hoa chat ma van dam bao diJ nong dp DNA cung nhu* d6 tinh sach cua DNA de tien hanh phan l>ng PCR, gop p h i n giam chi phi gia thanh cua xet nghiem.
V i e t n a m J o u r n a l of Physiology 2 1 ( 2 ) , 6 / 2 0 1 7 I S S N : 1 8 5 9 - 2 3 7 6
3.2. T I le phat hidn da hinh gen CYP2C19 va ITGB3 b i i n g kT t h u g t
Realtime - PCR
Bang 3.2. Ty Id phdt hien da hinh gen CYP2C19 va ITGB3 a nhdm nghidn ciru Chi so
Gioi Nam
NO Chung
Gen CYP2C19
Da hinh CYP2C19*2 B a hinh CYP2C19'3 16(47,1%) 1(2,5%)
2 (33,3%) 2 (33,3%) 21 (52,5%)
Gen ITGB3 Da hinh PLA1/PiA2
3 (8,8%) 1 (16,7%)
4 (10%) Nhan xet: Ket qua d' bang trdn cho
thdy, ti le cdc da hinh gen CYP2C19*2 vd CYP2C19*3 if nhdm nghidn c u u Id t u a n g d l i Idn: cd 52,5% benh nhan mang it nhat 1 alen CYP2C19 lam giam chuc nang c h u y l n hda clopidogrel. Bdn canh dd cung cd 10% bdnh nhan
c6 da hinh gen ITGB3 lien quan den de khang aspinn.
3.3. So s a n h ket qud cua ky thuiit giai t r i n h t\y gen vd-i k i t qua cua ky t h u a t realtime-PCR
So sanh k i t qua cua 2 ky thudt dup'c trinh bay dud'i bang sau Bang 3.3. So sdnh kdt qua realtime - PCR vd'i giai trinh t y gen
IvlSu
1 2 3 4 5
K i t qua 2 ky thuat giong nhau
X X X X X
Ket qua 2 i<y thuat Ithac nhau rviiu
6 7 8 9 10
Ket qua 2 kJ thuat giong nhau
X X X X X
Ket qua 2 k f thu$t khde nhau
Nhan x4t: Ket qua xac dmh da hinh gen CYP2C19 vd ITGB3 d hai ky thugt la hoan toan g i l n g nhau.
3.4. TJ- Id cac da h i n h gen CYP2C19 va ITGB3 o* n h o m n g h i d n c i r u
Bang 3.4.
Gen
CYP2C19*2 (G681A) CYP2C19*3
(G636A) PIA1/PIA2 (T1565C)
Ket qua xac djnh kieu gen CYP2C19 va
Binh thuang
(%)
n = 2 2 (55%) n = 37 (92,5%) n = 36 (90%)
iTGB3 o Tan so k i i u gen Dj hop t u
(%)
n = 1 4 (35%) n = 3 (7,5%) n = 4 (10%)
' nhdm nghiSn dru
D i n g hpp tCp da hinh
(%)
n = 4 (10%)
n = 0 (0%) n = 0 (0%) Nhan xet: Ket qua tren cho thdy,
bdnh nhdn mang alen CYP2C19*2 chiem td'i 45%, chCi yeu cd k i l u gen dj hp'p t u
(35%), kieu gen dong hp'p t i l c h i l m 10%.
C d 7,5%. bdnh nhan mang alen CYP2C19*3, deu c6 k i l u gen dj hp'p ti>.
V i e t n a m J o u r n a l of Physiology 2 1 ( 2 ) . 6 / 2 0 1 7 I S S N : 1 8 5 9 - 2 3 7 6
Cd 10%) bdnh nhan mang alen PIA2 - d i u cd k i l u gen dj hp'p ti>.
4. BAN LUAN
Ngay nay, p h u a n g phap ARMS- Realtime- PCR ngay cang dup'c ap dyng r^ng rai d' n h i l u nud'c trdn the giai doi vd'i nhO'ng benh cd cdc ddt bien d i l m da dup'c b i l t trud'c. Phan i l n g nay dung cac m l i dac hieu cd trinh t y d i u 3' bo sung vdi alen dot bien vd mdt mdi chung ngup'c chieu vdi vd'i moi dac hieu alen. S y cd mat cua dot b i l n d u a c the hidn bang san pham ADN k h u l c h dgi vdi cdc kich t h u d c khac nhau da bidt trudc. E d t b i l n dup'c xac djnh k i l u gen bdng ARMS-realtime - PCR. N h u vay, de xac djnh d u a c dgt b i l n d i l m , Chung ta phdi t h i l t k l 3 mdi. Trong dd mdt moi d u a c SLP dung chung cho ca m l u binh t h u d n g va mau cd dot b i l n , cd t h i la moi xuoi hoac mdi ngup'c. Hai m6i cdn Igi cd trinh t y gidng nhau, t r u vi tri nucleotid d ddu tgn cung 3". Mdi binh t h u d n g cd trinh t y nucleotid d ddu tan cung 3' b l sung vdi trinh t u ADN binh t h u d n g . Moi dot b i l n cd nucleotid d dau tan CLing 3' bo sung vdi trinh t y ADN cd logi dot bien d i l m do. Theo nguydn tac nay, mdi binh t h u d n g chi khuech dai dogn ADN binh t h u d n g ma khong k h u l c h dai dogn ADN cd dot bien, va ngup'c lai moi dgt b i l n chi k h u l c h dai dogn ADN cd d$t b i l n ma khdng k h u l c h dgi doan ADN binh t h u d n g . Do dd, p h u a n g phdp nay chJ thanh cdng khi thiet k l dug'c m l i dgc hidu hoan toan vdi ADN dich d dau 3'. Sau khi t h u nghiem bd kit ARlVIS-realtime -PCR cua hang Lytech. Nga va so sdnh vdi k i t qua giai trinh t y gen, nhan thciy ky thudt ARMS-PCR khdng chi cho k i t qua chinh xac, md cdn rut ngan thdi gian xet nghidm, quy trinh ky thudt den gian. Bang cdch gidm the tich kit tdch DNA va the tich mau t u a n g u n g , chung tdi nhan thay vdn cd the dat nong do DNA vd dd tinh sach dam bao cho kT thuat ARMS - Realtime - PCR va giam dug'c gid thanh xet nghidm.
Benh mgch vanh ngay cang gia tang va chiem ti 1$ Idn trong cdc bdnh tim mach ndi ridng vd md hinh bpnh tat ndi chung.
Ligu phap c h I n g k i t tap t i l u cdu ddng vai trd t h i l t y l u trong d i l u tn hoi chung vanh cap vd cac thu thugt can thiep mgch vanh qua da. Hign nay, phuang phap d i l u tri phoi hg-p aspirin va clopidogrel d u g c xem nhu lieu phdp khdng t i l u cdu chuan cho cac benh li ndy. Tuy nhien, n h i l u nghidn c u u da chi ra s y d l khang vdi aspirin va clopidogrel, lam giam hidu qua dieu trj.
Theo nghien c u u cCia Bons T. vd cs (2009), ty le khdng dap ung vdi aspirin dao ddng t u 5 - 60%), vdi clopidogrel 6 - 25% va ca hai thudc Id 10,4% [6]. & V\ei Nam, D 5 Quang Hudn (2013) khi khdo sat 174 bdnh nhan mach vanh t h i y ty Id khdng dap ung vdi aspinn Id 21.3%. vdi clopidogrel Id 26,4% [7]. Nhu vay, nghidn c u u ve cac y l u td lidn quan den s y d l khdng clopidogrel vd aspinn, ddc bidt yeu td di truyen Id rdt can t h i l t trong vide phdng va d i l u tri benh mgch vanh.
Trong nghien CLFU cua chiing tdi, cd 52,5% sd bgnh nhan cd mang it nhdt 1 alen CYP2C19 lam gidm kha nang c h u y l n hda clopidogrel (CYP2C19*2 hoac CYP2C19*3). K i t qua ndy t u a n g dong vdi nghien cu-u cua Liu Mao va cs (2013): cd khoang 55% ngudi chau A mang it nhdt 1 alen lam giam hogt tinh cua enzym CYP2C19 [8]. Trong dd, chu yeu la cac bgnh nhan cd k i l u gen dj hgp CYP2C19*2 (14 benh nhdn - 35%) va 3 bdnh nhan mang kieu gen dj hgp CYP2C19'3 (7,5%).
Nhu vgy, k i l u gen chuyen hda d muc trung binh (intermediate metabolizers) clopidogrel c h i l m ty Id t u a n g doi Idn. Kieu gen c h u y l n hda kem (poor metabolizers) - d i n g hgp CYP2C19''2 vd d i n g hap CY2C19*3 cd 4 bdnh nhdn ( c h i l m ti Id 10%. sd benh nhan nghien cuu). Theo k h u y i n cdo cua Hdi d u g c li lam sang vd dieu trj Hoa Ky (ASCPT) nam 2013 v l sCi' dung thuoc chong ngung tdp tieu c l u d y a
V i e t n a m Journal o f Physiology 2 1 ( 2 ) , 6 / 2 0 1 7 I S S N : 1 8 5 9 - 2 3 7 6
trdn k i l u gen CYP2C19 [9], thi cdc BN cd k i l u gen CYP2C19 chuyen hda trung gian hogc c h u y l n hda kem can xem xdt tang lieu clopidogrel hoac didu trj thay t h i prasugrel. ticagrelor...
Ngoai ra, nghidn c u u cua chung tdi cdn ghi nhan 4 bdnh nhdn mang da hinh Leu33Pro gen ITGB3 ( c h i l m 10% s6 bdnh nhdn nghien cilu), t i t ca cac bgnh nhan dd d i u cd kieu gen dj hgp t u PIA1/PIA2.
Khdng cd benh nhan mang k i l u gen dong hgp t u PIA2/PIA2. Theo nghien c u u cua Sperr W . R. va cs (1998) tren ngudi Chau Au cho thay 20 - 30% nhdm nghidn c i l u mang k i l u gen PIA1/PIA2 vd 1 - 3% mang k i l u gen PiA2/PIA2 [10]. Nghien c u u nay cung chi ra r§ng kieu gen PIA1/PfA2 va PIA2/PIA2 ldm tdng phdn ung hoat hda thrombin han so vdi k i l u gen dong hgp t u PIA1/PIA1, do do lam gidm tdc dyng chong kdt tap t i l u c l u cLia aspirin, hay ndi cdch khac, ngudi cd mang alen PIA2 se tang kha nang khdng thudc aspirin h a n . Nghien c u u cua Jussi M. vd cs (2000) d ngudi Phan Lan cho thdy trong nhdm nghien c u u cd 7 3 , 1 % mang k i l u gen PIA1/PIA1, 24,2% PIA1/PIA2 vd 2.7%
PIA2/PIA2 [11]. Nghien c u u cung chi ra mdi lien quan chgt che giu'a alen PIA2 va s y gia tang cdc bien co tim mgch nhu h u y l t khoi ddng mach hoac dot quy.
Vdi nhO'ng ket qua nghien cuu trdn cua chung tdi, b u d c d i u cho thay ti lg mang da hinh gen CYP2C19 lam giam kha nang chuyen hda clopidogrel vd da hinh gen ITGB3 ldm giam dap ilng vdi aspirin d nhdm bdnh nhan mach vanh Vidt Nam la tuang doi Idn, can cd s y quan tdm can t h i l t t u phia cac bac sT lam sang t r u d c khi dieu trj phac do chong k i t tap tieu cdu kep.
5. K^T LUAN
Ti Id bgnh nhan mang da hinh gen CYP2C19 *2 va CYP2C19*3 d nghidn c u u Id 52.5%, ti Id bgnh nhan mang da hinh Leu33Pro gen ITGB3 la 10%. Nhu vay sd
bdnh nhdn cd kha nang ddp ilng vdi clopidogrel va aspirin kem la tuang d l i Idn, can cd s y xac djnh kieu gen d l can nhac l i l u lugng thuoc, hoac thay t h i thulc cho phu hgp trong phac dd d i l u trj cua tilng benh nhan sau dgt stent DMV.
Ky thugt ARMS - realtime - PCR la phugng phdp xac djnh da hinh gen nhanh chdng, chinh xdc va chi phi phCi hgp vdi dieu kien Vidt Nam, cd kha nang ilng dung tot trong lam sang.
6. KI^N NGHI
Qua k i t qua nghidn c i l u budc d i u v l hoan chinh va ung dyng ky thudt ARMS - realtime - PCR d l xac djnh da hinh gen CYP2C19 va ITGB3 d benh nhan mach vdnh, nhan thdy can dua xet nghidm nay trd thanh t h u d n g quy d l xdc dinh k i l u gen lien quan d i n c h u y l n hda thulc clopidogrel va aspirin t r u d c khi d i l u trj cho nhdm bdnh nhan nay.
LOT cam o n : Nghien c i l u d u g c thyc hidn tgi trudng Dai hpc Y Ha Ndi, trong thdi gian tin thdng 6/2016 - thang 2/2017, vdi s y ho t r g cua cac dong nghiep tgi Bg mdn Y sinh hgc - Di t r u y l n .
TAI Lieu THAM K H A O 1 . Rosamond W, Flegal K, Furle K et al.
(2008) Heart disease and stroke statistics- -2008 update: a report from the Amencan Heart Association Statistics Committee and Stroke Statistics Subcommittee. Circulation ;117: e25-146.
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ARMS - REALTIME - PCR APPLICATION FOR DETERMINE POLYMORPHISM OF C Y P 2 C i g AND ITGB3 GENE IN CLINICAL DIAGNOSTIC CLOPIDOGREL AND
ASPIRIN RESISTANCE IN PATIENTS PLACED CORONARY STENT Nguyen Thi Minh NgocSTrieu Tien Sang^ Nguyen Thi Trang^
^Ha Noi Medical University
^Vietnam Military Medical University Coronary artery disease (CAD) is the most common type of heart disease. If is the leading cause of death In the World In both men and women. Numerous studies have shown that the polymorphisms of CYP2C19*2, CYP2C19''3 gene CYP2C19 and PIA1/P1A2 (LeuSSPro) of the ITGB3 gene are associated with clopidogrel and aspirin resistance. DNA was extracted from 40 patients. Using ARMS - realtime - PCR technique to Identify polymorphisms CYP2C19*2, CYP2C19*3 gene CYP2C19 and PIA1/PIA2 (Leu33Pro) gene 1TGB3. Results: In 40 samples, carriers of at least one CYP2C19 loss- of-functlon approximately 52,5% of the study population. CYP2C19*2: 10% homozygous genotype, 35% heterozygous genotype and 55% wild type. CYP2C19'3: 7,5%
heterozygous genotype and 92.5% wild type. Carriers of the Leu33Pro polymorphism of ITGB3 gene approximately 10% of the study population. Conclusions: Initial description of polymorphisms of CYP2C19*2, CYP2C19*3 gene CYP2C19 and PIA1/PIA2 (Leu33Pro) gene ITGB3 have been found to be high rate in patient group used clopidogrel and aspirin dual therapy. In this article, we describe commonly used ARMS- realtime - PCR as well as the application of these technology to the genetic diagnosis of resistance aspirin and clopidogrel.
Keywords: coronary artery disease, CYP2C19, cbpidogrel, ITGB3, aspitin resistance.
