PHAT HIEN DOT BIEN GEN G A Y B E N H BETA - THALASSEMIA BANG KY THUAT MULTIPLEX ARMS - PCR
Tran Van K h a n h \ Pham Thanh L o a n \ Ho Cam T u \ Nguyen Du>c H i n h \ Tran Thj Oanh^ Tran Huy T h | n h \ Ta Thanh Van^
^Trwdng Dai hpc YHi Ndi, ^Cpc Khoa hoe Cdng nghe vi Dao tao, Bd Yti Beta - thalassemia (fi - thalassemia) li bdnh di truyen lin nhiem sic thi thwdng gay do giim hoac mat hodn toan sw ting h^p ehu6i globin p. Cho din nay khoang han 200 dot biin phat hidn trdn gen p - globin dwp^ chirng minh la nguydn nhdn gay bdnh jS - thalassemia. Nghidn cwu dwac thwe hidn nhim xic dinh mdt s6 dot bien phi biin trdn gen j8 - globin d bdnh nhin p - thalassemia bing ky thugt Multiplex ARMS - PCR 52 benh nhdn (i - thalassemia dwgv lira chpn vio nghien ciru de xac dinh ddt bien. Kit gua cho thiy 39/52 trwdng hgp mang dot bien chiim ty Id 75% vdi 9 loai dot bien pho biin trdn gen gen p - globin. Dot bien Codon 41/42 (- TCTT) va dot biin Codon 17 (A > T) ehiim ty Id cao nhit la 30% vi 21%.
Keywords: Beta - thalassemia. Multiplex ARMS - PCR
I. DAT VAN Bt
Beta - thalassemia (|3 - thalassemia) Id bgnh di truyen ldn nhiem sac the thudng do ddt b i i n gen p - globin (gen HBB: Haemoglo- bin beta gene Id gen tong hap chudi p cua phan t u hemoglobin), gdy giam hoac mat hodn todn s y t i n g hap chudi globin p, Ddy Id mpt trong nhdng benh huyet sdc to p h i bien nhat, phdn bo trdn todn the gidi cung nhu d Vidt Nam vd khu v y c ehdu A [ 1 ; 2]. Theo thong kd eiia To chuc Y te The gidi (1981) ed khoang 241 tridu ngudi tren the gidi mac bdnh.
Cung theo u d c tinh cua T l ehue Y td T h i gidi (2008) hdng nam cd khoang 300,000 tre mdi de bj cdc t h i nang cua thalassemia Cl Vidt Nam, theo Nguydn Cdng Khanh, bdnh p - thalassemia Id nguydn nhdn hdng dau gay thieu mdu, tan mdu ndng d tre em [2]. Cho d i n nay, cd khodng han 200 ddt b i i n da d u a c tim thay trdn gen p - globin [1]. 6" Viet Nam, mdt
o/a chi lidn h$: Trin Van Khinh. Tmng tim Gen - Pmtein, Tnrdng B^i hoc Y Hi NOi
Email: vankhanh73md(Si.vahoo.com Ngiy nhin: 04/8/2014
Ngiy du^ chip thuin-16/9/2014
SO nghidn cuu da cdng b6 ed 8 dot b i i n gdy ra 95% ede trudng hp'p p - thalassemia gom Cd17 (AAG - TAG), Cd41/42 (- TCTT), -28 (A
> G), Cd71/72 (+ A), IVS1 - 1 (G > T), IVS1 - 5 (G > C), IVS2 - 654 (C > T) Cd95 (+ A) vd ddt b i i n Cd26 (GAG > AAG) gdy the p - thalassemia k i t hap vdi benh h u y i t sac to E (HbE) [3]. p - thalassemia Id bgnh thieu mdu tan mdu nang va chua ed phuang phdp dieu trj thgt de. p - thalassemia t h i ndng vd t h i trung binh thudng bleu hien thieu mdu muc dd tCr trung binh den nang, cupc song phy thupe vdo viec t r u y i n mdu va thai sdt sudt d d i , n g y d i bgnh cd c h i t luang cude song thap dong thdi eiing Id gdnh ndng cho gia dinh va xa hdi. Xac dinh dot bien gen tren benh nhdn d l tien tdi phdt hien ngudi Idnh mang gen benh vd chan dodn trude sinh de ed nhung t u vdn di truyen thich hap ddng vai trd quan trpng gitip ngdn ngda va Idm giam ty Id mac bgnh. Ky thuat Multiplex ARMS - PCR da d u a c sir dung kha rpng rai de phat hidn ddt bien gen gay benh p - thalassemia do cd dd chinh xde eao, gid thdnh re vd it phdc tap [4;
5; 6]. 6/ Viet Nam vide dp dung cdc ky thugt
TCNCYH 90 (5) - 2014
smh hpc phdn t u tai cac tabo khac nhau de xac djnh dot b i i n gen eac benh ly di truydn ngay cdng dupe k h u y i n khich m d rpng d i giiip ich cho cdng tdc chan dodn benh. Xudt phdt t u t h y c t i tren, nghidn c y u d u o c t i i n hdnh vdi muc tieu: Xac dinh mdt sd dot b i i n phd b i i n tren gen 3 - globin d benh nhan 3 - thalassemia bang ky thuat Multiplex ARMS - P C R .
II. 061 TygNG vA PHU'aNG PHAP
1. Doi tu'ang
52 benh nhan duae ehan doan mdc benh 3 - thalassemia tai khoa Huyet hoc Truydn mau, benh vien Bach Mai dya vao trieu chung lam sang va can Idm sang dien hinh, 5 mdu DNA ngudi binh thudng d u a c s u dung lam mau doi chung.
2. Phu'ang phap 2.1. Ky thuat tach chiet DNA + DNA du'p'c tdch c h i i t t y mau ngogi vi eiia bdnh nhan p - thalassemia, s u dung QIAGEN Kit.
+ K i l m tra ndng dd vd dp tinh sach eiia DNA d y a c tdch chiet bdng phucng phdp do quang trdn mdy NanoDrop: nong dp DNA 300- 380ng/ml; danh gid dp tinh sach bdng ty 1^
A2S0nrr/A280nm = 1 , 8 - 2 , 0 .
2.2. Ky thuat Multiplex ARMS - PCR Multiplex ARMS - PCR d y a c dung de sang Ipc s y hidn dien eCia nhieu alen dot bien. Moi phan y n g se diing n h i i u moi ddc higu cd trinh t y dau 3' bo sung vdi alen dot bien vd mdt moi Chung ngupe c h i i u vdi m l i dac hidu alen, S y hien dien cua cae alen dot b i i n d y y c bieu hien bdng san pham DNA k h u i c h dai vdi cac kieh thude khac nhau da biet trude (hinh 1).
Moi t h i i t k i s u dung de phdt hidn 8 ddt bien gdy benh p - thalassemia vd dot b i l n gay benh HbE pho bien d Viet Nam bao gim Codon 41/42 (-TTCT), Codon 17 (A > T), Codon 71/72 (+ A), IVS I - 1(G > T), nt - 28 (A
> G), IVSII - 654 (C > T), IVS I - 5 (G > C), Codon 95 (+ A) va Codon 26 (A > G) [ 1 ; 7; 8], Mdi npi chuan d u a e diing de kiem tra thao tac ky thuat vd chat l u a n g gen.
h" \fiM .
-28 bz^bgl i : : c d i 7 ; — [ • " ^ ' ' P — > - : C d 2 6
IVS M IVS 1-5 C d 4 i ; 4 2 Cd71/72 C d 9 5
l. 2 6 8 b B _ ' . ^ ^ ^ 3 4 0 b p .'
. 344bp J , ^ 0 6 b D
^ ^
^ ^ ^ ^
r r r
FR
IVS 11-654
: M a i C h u n g : M 6 i n a i chu4kn (P^J : M o i d a c h i f u
: M o i x u 5 i : M S i ngu>a>c Hinh 1. IWinh hpa cac moi va vj tri gan tren gen p - giobin
su> di^ng trong iVIuitipiexARMS - PCR
TCNCYH 90 [z.:
MOI chung B d u a c s u dung ciing mdi dac hidu dot b i l n IVS II - 654, moi chung H d u p e s u dung Cling moi dac higu dot b i i n Cd26. Cdc mdi dde hieu cdn Igi cd eiing mdi chung E.
2.3. Ky thuat ARMS - PCR
Sau khi xdc dinh duae eac dot b i i n bdng ky thudt Multiplex ARMS - PCR, cdc dot bien ndy se dupe xdc djnh kieu gen ddng hap t u hay dj hap t u bdng ky thuat ARMS - PCR. Trong nghien edu nay, ky thudt ARMS - PCR se chi tien hdnh vdi cac moi thudng, Sau Iuan nhiet, san pham ciia phan y n g ARMS - PCR se d u a c dien di cung san pham cua phan ung Multiplex ARMS - PCR trudc dd.
rfiHT
Hinh 1. Minh hoa k i t qua cua Ky thuat ARMS - PCR
K i l u gen dong hap tCr (DHT): G i i n g 1 va 2 chi eho 1 vach d mdi dot b i i n (DB), mdi thudng khdng cd san pham; Kieu gen dj hap tCr (DHT): g i i n g 3 vd 4 cd san pham d ca mdi dot b i i n vd moi thydng.
2.4. Ky thuat giai trinh tip gen San pham PCR cua benh nhdn t u a n g dng vdi cdc dot bien khde nhau eua gen p - globin duac kiem tra lai bang ky thudt giai trinh t y gen theo quy trinh t h y d n g quy ciia Trung tdm Nghien cdu Gen - Protein.
3. Dao du'c nghien cii'u
Cae doi t u a n g tham gia nghidn cuu Id hoan todn t y nguydn va ed q u y i n nit lui khi khdng mudn tham gia nghien cuu. Cac thdng
tin lien quan den benh nhdn d u a c dam bao bi mat.
III. KET QUA
Trong sd 52 benh nhan da phat hien d u a c 39/52 (75%) bdnh nhdn mang it nhdt mdt trong 8 dot bien pho bien gay bdnh 3 - thalassemia d u a c nghien cuu, 10 trudng hap trong so ndy cd kem theo dot bien Codon 26 (G > A) gay t h i HbE tren lam sang (bang 1).
TCNCYH 90 (5)-2014
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- f
^ • ^ ^
s a *
S 3'
&'^-ir
13
9 *
ii
SB BT
Cdl7
i ^ ^ il
I00pli_
Hinh 2. Kit qua xac d[nh kilu gen bang ARMS - PCR cac may p.44, p,43, p,50, p.45 Gieng 1: iuilOO: Ttiang ctiuSn DNA l<icii thirdc lOObp; BB: m6i dOt bien; BT: m6i blnh thuong;
Gilng 2 va 3: Miu p.44 Dong hp'p tO Cd17 (A > T); Gilng 4 va 5: IVIiu (3.48 di ho'p ti> i<ep 2 d6t biln: Cd 71/72 (+ A) v4 -28 (A > G); Gilng 6 va 7: M5u 3.60 di hp'p t i i<ep 2 dot biln: Cd 41/42 (-TCTT) va -28 (A > G); Gieng 8, 9: Miu |3.45 di hop t i i<ep 2 dot biln: Cd 41/42 (-TCTT) va Cd17 (A > T); cung chirng ngi chuan ngu-cyi binh thu'O'ng 494bp.
Ket qua x^c dinh d6t biln gen da 6\jgc i<ilm tra iai blng kit qua gi^i trinh ty gen.
IVS 1-1 (G>T) c,-28A>G (g, 70516A>G
Hinh 3, Hinh anh dai dien ket qua gicii trinh ttF gen cua 2 dot bien IVS - 1 (G > T) vajUB (A > G)
Kit qua giai trinh tu' cho thay hoan toan phCi hpp voi l<lt qua sang lgc dot biln blng l<y thuat Multiplex ARMS - PCR.
TCNCYH 90(5)-2014
Bang 1. Cdc loai dot bien vd kieu gen gay benh p - thalassemia
Dang 6qt bien Kieu gen So lyang
Cd41/42(-TCTT) P°//p
Cd17(A>T) p°//p
Dj hp'p tu dan
(n = 17) Cd71/72(+A) P°//p
IVS I - 1 (G > T) P°//|3
IVS I - 5 (G > C) r//p
Dong hgp tCr (n = 5)
Cd41/42(- TCTT) // Cd41/42(- TCTT i°//3°
Cd17(A>T)//Cd17(A>T) i"//3°
IVS II - 654 (C > T)// IVS II - 664 (C > T) r//r
Cd41/42(-TCTT) // Cd17(A > T) Cd41/42(- TCTT)// Cd71/72 (+ A) p - thai//
p - thai (n = 7)
Cd41/42(-TCTT)// -28 (A > G) Cd17(A>T)//-28(A>G) Dl
hgp
tLP
kep (n = 17)
Cd71/72(+A)//-28(A>G) Cd17(A>T)//IVSI-1 (G>T) Cd41/42(-TCTT) // Cd26{G > A) HbE
Cd17(A > T) // Cd26(G > A) HbE P-thal //
HbE (n = 10)
Cd71/72 (+A) // Cd26 (G > A) HbE IVS I -1 (G > T) // Cd26(G > A) HbE
IVS 1-6 (G > C) // Cd26(G > A) HbE Cd96 (+A) //Cd26(G > A) HbE
i"//p°
i"//p"
3°//3*
3°//3*
3°//3*
3°//3°
37/ HbE P°// HbE 37/ HbE P7/ HbE 3* // HbE 3°// HbE B?nh nhan Co dot bien
Bgnh nhan khong phat hien thay dot bien Tong
Trong so 39 b#nh nhan ph^t hi6n du'gc dot biln, cd 66 alen dot biln gom: 17 alen dot bien Cd 41/42 (-TCTT) Chilm 30%, la dgt biln pho biln nhit. 12 alen dot biln Cd 17(A > T) chiem ty Ig cao thij hai vol 21%. Hal loai dot biln Cd 95 (+A) va IVS II - 654 (C > T) chilm ty le 2%. 10 alen dot bien Cd26 (G > A) gay HbE chiem 18% (bleu dfl 2).
TCNCYH 90 (5) - 2014
^ U Y H Q ^
Cd26(G>A) HbE18%
Cd41/42(-TrCr)
30% DCd41/42{-rrCD
DCdn(A>T) D 28 (A>G) ! D Cd71/72(+A) j
• IVS I-I(G.T) B IVS I-5(G>C)
• Cd95(+A) i • IVS II-654(OT)
D Cd26(G>A) HbE ;
Bieu do 2. Ty le eac dang dot bien du'p'c phat hien
Trong so 39 trud'ng hop vdi 56 alen dot b i l n da phdt hien cd 17 trud'ng hap (43,6%) mang k i i u gen di hap t u vai mdt dot bien; 5 trud'ng hap (12,8%) mang k i i u gen ddng hap t u ; 17 trud'ng hap (43,6%) mang kieu gen dj hap t u kep vai 2 dot b i l n (trong do 10 trud'ng hap cd kieu gen di hap t u p - thalassemia k i t hap HbE). Tdt ea 10 trud'ng hap dot b i i n Cd26 (A > G) gdy t h i HbE ddu ed kieu gen dj hap t u
Dlho^ttukepj]
thai, 17 (43,6%)
fionghof tu-iS, (12,8%)
• Bong hop t«
• Dihoptidim DDihoptakepp-M
Di h(rp tu doD, 17, (43,6%)
Bieu do 3. Ty Id kieu gen phdthien trong nghien CLPU
TCNCYH 90 (5)-2014
Bang 2. Ty le cac dot b i i n phat hjen du'ac so sanh vd'i nghien cu'u khac
Bu'd'c sang
loe
Dot bien Kieu hinh
Nghien CIFU N . K . Han- Anchalee Winichagoon nay n = 52 Hoan [46] [1] n = 252 [47] n = 375
(%) n = 3 3 ( % ) (%) (%)
Cd 41/42 (-TTCT)
C d 1 7 ( A > T ) Cd71/72(+ A) 1 IVS 1-1 (G >T)
-28 (A > G) IVS 1-6 (G > C) Cd95(+A)
^ I V S I I - 6 5 4 ( C > T ) 3 Cd26 (A > G)
Dot bien khac
3°
3°
3°
3*
3°
3*
3°
3*
HbE
17(30,4%)
12(21,4%) 6 (8,9%) 6 (8,9%) 3 (5,4%) 2 (3,6%) 1(1,8%) 1 (1,8%) 10(17,8%)
11 (33,3%)
4(12,1%,) 4(12,1%,) 3 (9%)
0 0 0 4(12,1%) 7 (21,2%)
-
107 (42,5%) 46 (7,9%)
8 (3,2%) 8 (3,2%) 28(11,1%) 11 (4,4%)
0 19(7,5%)
24 (7,6%)
156(41,6%,)
62(16,5%) 8(2,1%) 5(1,3%) 34(9,1%) 16(4,3%) 1 (0,3%) 28 (7,5%)
65(17,3%)
{-) • cac dot bien khong dirge sang lgc trong nghien ciru.
IV. BAN LUAN
Ky thudt Multiplex ARMS - PCR d u a c s u dung de phdt hidn 8 loai dot b i i n pho b i i n vd dot bien Cd26 (A > G) gdy t h i p-thalassemia k i t hap vd'i HbE, la nguydn nhdn gay 95%i benh p-thaiassemia d' Vidt Nam [3]. Nhdm nghien eiru da s u dyng ky thuat Multiplex ARMS - PCR va da phat hien dwgc 39/52 trud'ng hap mang dot bien ehiim 75%i. 13/52 trudng hap (ehiem 25%) khdng phdt hidn dot bien ed the do nhung dot bien khac khdng ndm trong so eac dpt bien sdng Ipe trong nghidn etru ndy, can lam thdm ede phuang phdp khde d i xde djnh dot bien. K i t qua nay thap han so vai nghien euu eua Nguydn Khde Hdn Hoan tai benh vien TCr DO, khi ty Ip phdt hidn dot bien trong bdo cao ndy la 100% (vd'i n = 27) [9].
Tat ea 8 loai dot bien gdy benh P-thalassemia vd dot b i i n Cd 26 (A > G) d i u dwgc phdt hien trong nghidn euu. K i t qua nay phu hpp vd'i nghien cuu eua ML. Saovaros Svasti tren qudn the ngud'i Viet Nam khi nghidn eiru vd ty Id xudt hien eua eae dot b i i n ke trdn vai ea mdu t u a n g d u a n g la 50 benh nhdn [3], ddng thd'i eung phu hap vdi k i t qua nghien ciru cua mdt vai tae gia khde tren ede qudn t h i ldn can Id Thdi Lan vd midn Nam Trung Quoe [1; 8]
Trong sd cae dot bien da phat hien, dot b i i n Cd41/42 (-TCTT) c h i i m ty Id cao nhat (30%), tiep theo Id dot bien Cd17 (A > T) (21%), dot bidn Cd26 (G > A) HbE (18%), ty Id nay t u o n g irng vd'i cae nghidn eiru da d u p e bao cao va cac tdc gia khde [1; 9; 10; 11].
T y le eua dot b i i n C d i 7 (A > T) trong nghien ciru la 2 1 % , eao han so vdi nghien
TCNCYH 90 (5) - 2014 23
cuu eua Nguydn Khde Hdn Hoan (12,1%), Tuy nhidn, ty Id eua dot b i i n Cd 71/72(+A) Id 9%
ed phan thdp han (12,1%) [9], TJ Id ede dpt b i i n - 28 (A > G), IVS I - 1 (G > T) cung cd s u khde biet. Trong bao cdo eua N g u y i n Khde Hdn Hoan dot b i i n -28 (A > G) khdng xuat hidn trong khi trong nghidn eiru eua chiing toi x u i t hidn 3 alen (ehiem 5%), s u khde biet nay ed the la do cd' mdu nghien cuu vd phdn vung dja ly, 00" m i u cua ehung tdi Id 52 vd nghien ciru tren cac bdnh nhan sinh song tai cac tinh phia Bae, frong khi ed" mdu eua Nguyen Khac Hdn Hoan la 27 vd nghidn eiru tai ede tinh khu vuc mien Nam, Vidt Nam.
Cdo dot b i i n edn l^i e h i i m ty Ip thap bao gom dot b i i n IVS I - 5 (G > C), IVS II - 654 (C > T), Cd 95(+A) lan luat Id 4%; 2% vd 2%.
Ty le nay eung phu hap vd'i nghidn eiru eCia D u a n g Bd True, Nguydn Khae Hdn Hoan va nghidn eiru eua Fllon D khi nghidn ciru tren ede doi t u a n g ed vCing sinh s i n g tai khu vuc phia Bde Vidt Nam [9; 12; 13],
Trong s6 39 bdnh nhdn phat hien mang dot b i i n trdn gen p - globin, phat hien d u a e 17/39 bdnh nhan (43,6%) mang kieu gen dj hap t u 1 ddt bien; 5/39 b^nh nhdn (12,8%) dong hap tir; 17/39 bdnh nhdn (43,6%) dj hap t u kep 2 dpt b i i n (trong dd 10 benh nhdn ed kieu gen p - thalassemia k i t hap HbE). K i t qua nay ed phan khac biet so vd'i kdt qua nghidn eiru cua ML. Saovaros svasti vd Anehalee hau het ede dpt b i i n tim d u a c trong cac nghien eiru trdn t6n tai a k i i u gen dong hap t u hay dj hap t u kep vai hai loai dpt bidn [ 1 ; 3]. Dieu ndy du'ac giai thich bd'i s y khac biet vd dd tuoi eua bdnh nhdn trong quan the nghien ciru: Cdc benh nhdn trong nghidn eiru trdn ehu yeu la nhOng bdnh nhi ed dp tuoi nho tCr 1 den 15 tuoi va da eo bieu hipn thidu mdu nang, phai truyen mau dinh ky (nhung doi t u a n g nay thud'ng mang kieu gen dong h a p tCr hoac dj
h a p t u kep 2 dpt b i i n ) , trong khi nchien ciru ciia ehOng toi t i i n hdnh tren nhi>ng b?nh nhdn cd dp tuoi chu yeu trdn 16 tu6i (41/52 trud'ng h a p tir 16 den 57 tuoi, ehi co 11 trud'ng h a p ed dp tuoi tir 4 d i n 16).
V. K^T LUAN
Nghien ciru da phat hidn d u a c 39/52 trud'ng hap mang ddt bien ehiem 75%i vai 9 loai dot b i i n trong d d :
Dot b i i n Codon 41/42 (-TCTT) va ddt bien Codon 17 (A > T) chiem ty Id eao nhat Id 30%
vd 2 1 % . Ddt b i i n Codon 95 (+A) va ddt bien IVS II - 654 (C > T) cd ty Id th^p nhat 2%. Ddt bien Codon 26(G > A) gay p - thalassemia ket hp'p vd'i HbE gap vd'i ty Id 18%».
Trong so 39 bpnh nhan phat hidn mang dpt bien tren gen p - globin phdt hipn d u a c 43,6%
mang k i i u gen dj hp'p t u 1 ddt b i i n ; 12,8%
dong hap tCr; 43,6%» dj h a p tir kep 2 dot bien (trong dd 10 mdu ed kieu gen p - thalassemia k i t hap HbE).
Lai cam cyn
De tdi d u a e t h y c hien vd'i s u ho trp' kinh phi cua d i tdi nhdnh edp nhd nu'd'c "Nghien ciru mpt so ehi so sinh hpe, trinh t u gen ty t h i ngud'i Viet Nam truang thanh vd dpt b i i n gen gdy bdnh thalassemia" thude de tai nhiem vu Quy gen "Danh gid dde diem di truyen ngudi Vidt Nam".
TAILI
IAM K H A O 1. -Anefialee Thedsawad, Sumalee jin- dadamrongwech, Suporn Chuncharunee, Punnee Butthep (2012). Multiplex ARMS- PCR Analysis for Nineteen p - Thalassemia Mutation, Journal of Hematology and Tranfu- sion Medicine, 22(1), 1 - 38.2. Nguyen Cong Khanh (1993). Tan suat bpnh hemoglobin a Viet Nam. Y hoc Vigt Nam, 174(8), 11 - 1 6 .
TCNCYH 90 (5)-2014
3. M.L. Saovaros Svasti, Tran Minh Hieu, Thongperm munkongdee, Pranee Winichagoon et al (2002). Molecular analysis of p-thalassemia in South Vietnam. American Journal of Hematology, 7 1 , 85 - 88.
4. Newton, C.R (1989). Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucleic Acids Res, 17(7), 2503 - 2516.
5. Old, J.M. (2003). Screening and genetic diagnosis of haemoglobin disorders. Blood Rev, 17(1), 4 3 - 5 3 .
6. Du'ang Ba T r y c (1997). Mpt sd kieu gen beta-Thalassemia 6' ngud'i mien Bac. Hdi nghi chuyen dS nhOng tiSn bd m&i ve Thalas- semia (Ban djch cCia D u a n g Ba Tn^c), 36 - 42.
• Urvashi Bhardwaj, Yao-Hua Zhang, Fred Lorey et al (2005). Molecular Genetic Confirmatory from New/born Southeast Asian- Screening Samples for the Common African- American, Asian Indian, and Chinese p- thalassemia Mutations. American journal of Hematology, 78, 249.
8. Zhang JZ, Cai SP, He X, Lin X (1998).
Molecular basis of beta-thalassemia in South China. Strategy for DNA analysis. Hum Genet.
78, 37- 40.
9. N g u y i n Khac Hdn Hoan, Pham Thj Vi^t Thanh, Tru>ang Dinh Kidt et al (2011).
Ch^n doan tru'd'C sinh benh thalassemia tren 290 truang hap. Tap chi Nghien ciru Y hgc, 74(3), 1 - 7.
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Summary
MOLECULAR DIAGNOSIS OF BETA - THALASSEMIA BY MULTIPLEX ARMS • PCR
Beta-thalassemia (P thalassemia) is an autosomal recessive blood disorder w/hich is characterized by an absence or a reduction in p - globin chain synthesis. More than 200 different mutations located within the P - globin gene are currently recognized throughout the world as causes for P-thalassemia. The aim of this study Is to detect the common mutations in p - globin using Multiplex ARMS - PCR. 50 unrelated patients with p-thalassemia were selected for this study. The results showed that 39 out of 52 patients having mutation, representing 7 5 % with 9 common mutations in the p - globin gene. The most frequent mutation was Codon 41/42 (-TCTT) at 30% and Codon 17 (A > T) at 2 1 % .
Keywords: Beta-thalassemia, Multiplex ARMS - PCR
TCNCYH 90 (5) - 2014