CHUYEN DE: HiNH THAI H9C lrNG Dl}NG PHAN rtcn ADN PHDI THAI Tty DO DE SANG LQC DI TRUYEN TRlf6'C SINH Tri~u Ti~n Sang*, Tr5n van Khoa*, Dinh Doim Long**, Nguy~n Duy Bic"" hqp nhorn mau Rh thai nhi, tntong hqp nhom rnau thai nhi Rr( -), 19i 10 tru'on\; TOM TAT Ma dau: Vi~c phat hi~n su ton tal cua ADN hop nhorn rnau Rh (+) phoi thai tlf va te beo phoi thai tlf huyet tu'dng cua me da rna mQt hu'ong chan SUMMARY doan tru'oc sinh moi b~lng bi~n phap khong can APPLICATION OF CELL FREE thi~p Ban chat cua vi~c chan doan khong xam FET AL DNA ANALYSIS ON Ian StI dl,!ngADN phoi thai luu hanh rnau PRENATAL GENETIC SCREENING ngoGlivi cua me de chan doen va sang IQcmQt so Introduction: The noninvasive nature of the b~nh di truyen Boi tu'Qng va phu'dng phep detection of fetal DNA in the maternal peripheral nghien ciru: 34 thai phu dang mang thai tU'tuan blood represents the greatest advantage over the thU' den tuan thU' cua thai kyoTrong do, 11 conventional methods of prenatal diagnosis The m~u cua gia dlnh mang gen b~nh tang san applications of this metrodology involve the thu'Qng th~n bam sinh, 10 m~u cua gia dlnh detection of the fetal sex, diagnosis, and mang gen b~nh Teo cd Duchenne du'QClay tU' evaluation of the prognosis of many diseases BGlihQCY Ha NQiva 13 m~u cua ngu'ai m~ mang Objects and methods: We selected the 34 nhom mau Rh(-) lay t9i trung tam Nghien cU'u pregnant women in the 8th week to the 9th wee;;' Sinh Y du'QchQc- HQcvi~n Quan y Be kiem tra of pregnancy including 11 samples of gene nhom mau Rh cua thai nhi, va sang IQc b~nh carrier parents of congenital adrenal hyperplasic tang san thu'Qngth~n va teo cd Duchenne chung (CAH), 10 samples gene carrier parents c; toi thu lay ADN du'QCtach chiet tU' huyet tu'dng Duchenne muscular dystrophy (DMD) obtained at cua m~ Ket qua: Boi voi sang IQCb~nh tang san the Hanoi Medical University and 13 samples of c thu'Qngth~n bam sinh co 5/11 tru'ong hQp khong mother with Rh (-) obtained at the Military phai dieu trl thuoc nua, tru'ong hQp tiep tl,lC Medical Academy To examine the performance dung thuoc den chQc oi de chan doan chinh of the Fetal RHD,CAH, DMD, we use tree xac thai nhi co mang gen CAH hay khong Sang circulating cell-free fetal DNA (ceff-DNA:; IQc b~nh Teo cd Duchenne co 4/10 tru'ong hQp extracted from maternal plasma Results: Fe!.'" am tinh v(1i gen SRY (khong mang b~nh), patients screened for' congenital adrer~~ 6/10 tru'ong hQp du'Qcchan doan (1tuan thU' 16 hyperplasia with 5/11 cases of non-medicatic·-' chQcoi thay tru'ong hQp bl b~nh va tru'ong treatment, cases continued dosing urr::, hQp du'dng tinh vdi gen SRY nhu'ng khong mang amniocentesis for fetal diagnostic accuracy w;:: gen DMD Chan doim du'Qcchinh xac 13 tru'ong CAH gene ,or not Screening for Duchen;-~ muscular dystrophy with 4/10 cases negative f~ HQc vi~n Qulin Y SRY gene (non-medical), and 6/10 cases we:; Khoa Y DuO'c- Dai.' Hoc Q1I6cGia Hit NQi diagnosed in the 16th week of amniocentes~,··· u~ * ** Chju tnich nhi~m chinh: TrAn Van Khoa Email: tvkhoabio@gmail.com Dt: 01684552737 Ngay nh~n bai: 5110/2014 Ngay dlly~t bai: 16/10/2014 130 found casesof the disease and casesof Dt·D gene with positive SRYgene, but not DMD ger ; y HQC VI~T NAM THANG 11 • 56 ()~C BI~T/2014 Accuretely diagnosed 13 cases of Rh between mother and baby, including cases of fetal Rh jlood group (-) and the remaining 10 cases Rh ,+) Keywords: fetal DNA; RhD; CAH, DMD; non-interference v , I £>J;\T VAN DE Nam 1969, Walknowska va cong sir da phat hien str co m~t cua t~ bao phoi thai W mau ngoai vi cua ngiroi me Error! Reference source not found Day la phat hien vo cung quan mo mot huong m6i d~y tri~n vQng chdn do{m tnr&e sinh b~ng bien phap khong can thi~p Di tat bd~ sinh da va dang la m(>t vAn d~ :6n kh6~g chi d6i v&i nganh sim khoa rna L~U hut S\l' quan tam eua to an xa h(>i Hi~n ~v viee di~u tri cae di tat bdm sinh v~n i~~ ~ue'kho kh~ va phfre·tl;lp Tang san tuy~n :huqng th~n bdm sinh (gQi t~t la CAHCongenital Adrenal Hyperplasia) lit m(>t 10l;li t~nh di truy~n lien quan d~n r6i IOl;ln :Drmone cua tuyen thugng th~n (cortisol ~dosterone) B~nh khong ph6 bi~n vai sm1t m~c b~nh lrl 1112.000-15.000 tre em sinh hong cau cua mau thai nhi eo th~ lot qua thai vao co th~ me, mau cua me se xwlt hien khang the Rh- chong Rh+ l~n eo thai d~u tien lUQ11g khang the khang Rh+ rnau ella me it, nhirng a nhirng l~n eo thai sau IUQ11g khang th~ khang Rh+ mau ella ngiroi me tang len nhieu, neu thai nhi la Rh+, khang th~ se khuyech tan qua a thai sang gay ngung k~t h6ng cdu cua thai nhi Vi v~y, rftt d~ gay xay thai, de non, thai chet hru, V oi cac yell cau cftp thi~t tren chung toi ti~n hanh nghien ciru va chdn hoa quy trinh sang IQc va chftn doan cae b~nh di truy~n qua mau m~ Thai di~m chung toi l\ra chQn la a tudn Clla thai kY· II DOl TU'Q'NG, HOA CHAT VA PHU'ONG PHAp NGHIEN C(rU 2.1 DBi tU'C~)'Ognghien cU'u d nghien Clru Clla chung t6i, chlmg toi Iva ch9n thai di~m cac tu6i thai til' tll~n thu cua thai ky d cac gia dinh co mang gen bi benh CAH, DMD va gen RhD (-) Cac m§u d~Q'clAy 5ml maLl trong 6ng EDT A, D?i hQc y Hit NQi chUng tai thu tMp duQ'c 11 mau mau ngo?i vi eLla m~ a cae gia dinh c6 B~nh lo~m duong ca Duchenne (Duchenne mang gen b~nh Tang san thlrQ'flg th?n b§m ular dystrophy - DMD) lit m(>t sinh Cae mfiu teo cO' Duchenne chung t6i thu benh thAn kinh ca - di truy~n ph6 bi~n th~p auqc 10 m~Ll mall ngo{li vi ella ngtrai ~, "ai' ty l~ meri m~c Ja 113.500 tre trai de m~ mang thai a eac gia alnll ell mang gen teo Benh DMD co d~c tnmg la thmH hoa va ca Duchenne T~i HQc vi~n QWln Y chung -suy 'y~u ca m(>t each tuftn ti~n d~n d~n ta,n t6i thu dtrqc 13 m~u mall ngol;li vi Ctta cac -,:1 tu vong suy tim va b(>i nhi~m ph6i ph\1 nCr mang nhom mali Rh(:J man~ thai, a dAu hieu Him sang ctm b~nh duqc nh~n tuan thlr va thlr ,9 Cac mau nghlen CUll a giai dOl;lntre tu 2, d~n tu6i C~c b~n~: sang 19C b~nh DMD va CAH duqc chl;ly duae coi nhu mac the n~ng neu phm nhfm gen tren gen SRY va gen GAPDH lam xe Ian mAt kha nang tv di ll;litnr&c tu6i n6i chlrng Cac m~u ch~n an nhom mau Rh c(1a thai nhi se dui chlrng Tftt ca cac P~\l nfr la Rh+ ho~c Rh- N~u thai nhi Rh+ dugc ti~n hanh Itty 5ml mau tofm phan va trinh mang thai Ian dau m(>t hrgng nho A 131 CHUYEN £H~: HiNH THAI HQC Huy~t nrong sau 1y Him ti~n hanh l~y 200pl huy~l nrong tach chi~t bang bQ kit cua Quiagen ADN sau i.ach chi~t dircc hoc huy~t tirong 2.2 H6a chAt nghien ctru tan 50~t1H20 deion _ H6a chAt tach cbi~t ADN: Ethanol 70% Thirc hien phan irng nhan gen voi gen Ethanol 100%, Proteinase K Niroc cat, B(> SRY gen RhD va gcn GAPDH noi chimg kit Quiagen Cac mfill ADN sau tach xong a trer _ Hoa chAt cho peR: PCR Reaction Mix se duoc nhan gen bang ky thuat Nested PCR DNA Polymerase, Reverse primer, Forward (peR lbng) cho gen SR Y va nhan gen PCR primer, Probe, Nuac cftt klnr ion va v6 trung thucmg, cho g,en GAPDH Voi trinh tv m6; 2J PIIU'Ol1g phap nghien cuu cua gen nhu sau: h~' SRY' va GAPDH ang tnn W va kiIC h th iroc san p am cua gen Size bn Tn B'aug 1: B' Primer 5511C Loai rca Gen eTA NNN CGe AGA GGC Gee AT 239 ly tam 5000 vong/phllt phut rbi lAy SRY ngoai Nested SRY GAPDH GAPDH F R TAG NNN CCA CGC eTG eTC CGG CAT NNN GAG CGT CCC TGG CTT CTT NNN ACA GCC ACA TTT GTC CCCCACACACATGCACTTACC CCTAGTCCCAGGGCTTTGATT 198 57°C 98 6011C Chulrinh nhi¢1 ella phan {eng (.1tr(J"c thuc hiin thea bang Lite/Ii aiiy: Ballg 2: Bang ehu lrlnh nhl~t elm philn lrng nhan gen rn61 ngoal va m6i gen SRY va elm trinh nhiet nhan gen GAPDH Chu trinh nhi~t ch~y nhan gen SRY mbi ngoai Chu t1'i11hnhi~t eh?y nhfm gen SR Y mbi Chu t1'1nhnhi~t Ch~lYnhf111 gen GAPDH n"c nOc 7t'C 41'C 95(lC 941>C 60°(' 41'(' - 22 ehu ky ehu ky ehu ky elm ky en 4"C 95llC 94"C 57llC 1'2"(' ehu ky phllt phllt phllt phllt 20 phut 95"C 94"C 5511(' 7t'C - phllt phllt phut phllt 20 phllt 30 elm ky ehu ky ehu ky chu ky (/.1 phllt phllt I phllt 20 phllt 45 chu ky chu ky I ehu ky (I.' ' • A A Cae IIl?'U mu nhan XC n awe ,Mn di (ren gel Axar""c 2% vm 0' ~n (he 110 V 150mA c'" A v6"i thai gian 30 phttf TEE IX 132 Y HQC VI$T NAM THANG 11· ~ 'J , 1II.KETQUA VA BAN LU;N 3.1 K~t qua sang loc b~nh tang san thU'Qngth~n bim sinh Sau da chuan hoa diroc ky thuat tach ::hi~t ADN phoi thai W va k5' thuat PCR nhfm gen SR Y chung toi tien hanh thir nghi~m sang loc benh tang san thuong th~n bam sinh tnroc sinh tuftn tlnr cua thai ky a D6i voi cac truemg h9'P thai duong tinh vui gen SRY sau nhan PCR thi kheng phai dimg thu6c Dexamethason, cae b~nh ffinlt 1: Hinh anh so £)~C BI$T/2014 nhan am tinh voi gen SRY se duoc cac bac si tiep we eho dung thuoc Dexamethason d~n tuan tlur 16 cua thai ky d~ tranh tnrong hop nam hoa thai nhi mang gen b~nh CAH D~n twln cac bac si sieu am lai va choc 6i voi cac thai am tinh voi gen SRY chAn doan co bi benh CAH hay khong tiep we phac d6 di~u tri benh bang Dexamethason de de ntra hay dung thuoc khong m~c benh Chtmg t6i thu duQ'c k~t qua di~n di san phdm peR nlm sau: \!; Marker 100 Am: cht'mg tim mUle edt Duang:c!llcng dmmg CAHI, CAH2, CAH3, CAH-I, CAH5, CAH6, CAH7 CAHN, CAH9 (,AHIO,CAHIlI'1 mdll b?nh nhan ngh;en cdc C1CU ffinl, 2: Hinh anh nhun gen GAPDH cua cac m5u sang 19C b~nh CAH fE .\1; Marker lOO I; C,ic mJu l-llllrang Am: ch,rng am nwi< cdl Vilang: Ch'fn!{ dmmg t'mg vai mftu CAHI, CAH2, CAH3 CAH.J, CAH5 CA/-/6 CAH7 CAH8, t-" CAH9, CAH10,CAHl1 hI cae mftu h?nh nhan nghien etcu ~: t Ti< k~t qua di~n di chung t6i nh~n th5y rAngco 5111 truimg hgp duong tlnh viri gen SRY t va 6/11 trlrong hgp am tinh viri gen SRY T5t cil II truimg hgp dbu cho ktt qua duang ~ [ tinh v6i gen nQichung GAPDH t,I ! 133 CHUYEN DE: HiNH THAI HQC Biz MaBN Mii BN CAH7 AHI CAH8 CAH2 CAH9 CAH3 CAHIO 8 CAH4 CAH11 8 CAH5 CAH6 thai ky Cac gia dinh co ong ngoai ~ Til qua tren chung toi nh~n ding benh va truyen gen benh eho me thai nhi \i· 11 benh nhan sang lee tang san thuong thai nhi co kha nang mang gen b~r-~ th~n bam sinh co tnrong hQ'P duong diroc di truyen nr ong ngoai, D6i voi cac thzi tinh voi gen SR Y va khong phai dung thuoc nhi rna am tinh vci gen SRY se khong ~ nira, Con tnrong hQ'Pam tinh voi gen SR Y benh Con nhtmg thai nhi dirong tinh voi gea ti~p we phai dung thuoc d~n tu~n thir 16 cua SR Y se co th~ mang gen benh va bi b~nr~ thai kyoTheo tac gia Du, Zou Pan va cong Sl! Vi~c chan doan chinh xac thai nhi co man; nam 2009 da sang loc va chAn doan gen benh hay khong se duoc ti~n hanh choc cong cac tnrong hQ'P mang thai b] benh tang 6i va la xet nghi~m a tu~n thtr 16 eua thai k::_ san thlTgng th~ bAm sinh thong qua ADN Ch(mg toi l\ra chQn duqc 10 gia dinh co ng~:· phoi thai tl! mau ngo~i vi ella mtr· co sinh bi Teo co Duchenne dugc gui rj 3.2 K~t qua sang IQc b~nh Teo co' Trung tam Cong ngh~ cao - D~i hQc y H3 Duchenne NQi Ch(mg toi nhfm gen SR Y va ge:: Cae gia dinh, dong hQ co ti~n su sinh GAPDH nlm quy trinh tren v6i lOmb bi teo co Duchenne se dugc sang IQc b~nh nghien ClTll va thu duge k~t qua nhu sau: tru6c sinh b~ng ADN phoi thai t\f mau ngo~i vi Clta m~ a tuAl~thu clm M; Marker 100 Am: cht'rngam murc Dlumg: cln,nK dmmg DMDI, DMDl, DMD3, DMD4, DMD5, DMD6, DMD7, DMD8 DMDY, DMDIIJ, ki cae mall b?nh nht1n nghien cteu 134 y H9c VI~T NAMTHANG11 - 56 £)~c BI~T12014 Am: chimg am male Duong: chung duang DMDJ, DMD2, DMD3, DMD4, DMD5, DMD6, DMD7, DMDR, DMD9, DMDIO, III cue mJu \f: Marker 100 b?nh nhim nghien ciru Tir hinh di~n di san pham PCR nhan gen SRY d~ sang lee benh Teo co Duchenne mroc sinh chung t6i nh~n th~y r~ng cac mau len bang dep, TO net, chirng am kh6ng bi nhi~m, clurng dirong len bang t6t Co 4/10 mau am tinh voi gen SRY, nhimg gen nQi B·(lug_ 4: B'ang k"et qua sang , MaBN : : Tu3i thai SRY Sieu am thai 16 ) tuan QC «n chirng van duong chirng to chung t6i van tach duoc ADN huy~t nrong Con 6/10 mau duong tinh voi gen SRY va duong tinh voi ca gen nQi chirng GAPDH v~y kh6ng co m&unao duong tinh gift v6i gen SRY eo co Due henne Sieu Choc Ai chAn doan (+) (+) (+) IDMDOl (-) (+) (+) : DMD02 (+) (+) (+) i DMD03 (-) (+) (+) : DMD04 (-) (-) : DMDOS Qua bang s6 li~u tren chung t6i nh~n thay rang 10 tnrong hop co tnrong hQ'pdtrong tinh v6i gen SRY va truang hQ'p am tinh v6i gen SRY 86n trtremg hgp am tinh v6i gen SRY se kh6ng c~n ch9C 6i d~ ch:ln doan b~nh Teo co Duchenne nua Con truemg hgp duong tinh v6i gen SRY se duQ'cch9C 6i d~ ti~n himh xet nghi~m b~nh Teo co Duchenne t~i f)~i h9C Y Ha NQi va cho k~t qua 3/6 tnrang hgp mang b~nh Teo co Duchenne tnrang hQ'pkh6ng mang gen b~nh thai nhi sinh binh thuang MuBN Tu3i thai SRY ChQc I am 01 thai 16 tuan chAn doan ) (-) (+) (+) DMD06 (-) (-) DMD07 (-) (-) DMD08 (-) (-) DMD09 (+) (+) (+) DMDIO Tac gia Sekizawa, A.• T Kimura M Sasaki va cong sv (1996) da chan dofm c6ng B~nh teo co Duchenne tren cac m~u hfmg du co nhan Cllaph6i thai lUllhimh mall ngo~i vi Cllamy 3.3 K~t qua chan doan Slf bit dAng nhom mau Rh gill'a m~ va thai nhi Chlmg toi chAn do{mbitt dfmg nh6m mall gifra my va can thong qua ADN ph6i thai W mall ngo~i vi Clla my d6i v6i 13 tnrang hqp chAn doan va duQ'c chAn doan 13S CHUYEN DE: HiNH THAI H9C kiem tra chinh xac lai sau sinh Viec chan doan Rh sau sinh diroc thuc hien tai phong 107 Trung tam nghien ciru Sinh Y Primer Gen RhO: RhO-FSAAAGGGGTGGGT AGGGAA TA-3' RhD-R:5'_AGGTAGGGGCTGGACAGAA-3' San phflm phan irng cua gen RhO 10 Duoc - HQc Vi~n Quan Y 366bp ~ Chu trinh nhie: cua phan irnu: Buoc I chu ky 94"C - phLlt 94"C - I phllt 30 chu ky S4°C - phut 7211C- phut chu ky 72°C - 10 phut Buae chu ky 4C1C -vo , cung Buae4 San phdm peR sif duac kiem Ira bang kY Ihllal di tren gel agarase 2% Dim ili?n di TEE IX Chu trinh nhtin gen GAP DH ,1l((!cthuc hii n nhit chu tnnh d tren Son phdm peR cua ~en Buac2 men npi chzrn/( GAPDH Iii 98bp Sail nhtin gen GAPDH v,i gen RhD ad chan ,Iotin nhom mJ, Rh cua thai nh! chung t6i thu duoc kdlljl/(i SCIlI: Hinlt Hlnh anh di~n di ki~m tra nh6m mall Rh Clla thai nhi M am dVD'ns rhl 1hZ rhl rh4 IhS rn6 rit7 ,hi 1M ,hlO rhll rh l~ Itln }G(.hp IO!U M; Marker 100 Am: c:hll'11g am ntf(5-c cdr Dmmg: c:hl'mgcheong Rhl, Rh2, Rh3, Rh4, Rh5, Rho, Rh7, R.h8,Rh9, Rhl0, Rhll, Rh12, Rh13 hi cue mJu h~nh nhan nghWn cleu Bang Bang Mil b~nh nhan RHOI Rh02 RH03 RH04 RHOS RH06 RH07 136 Tu8i thai h'1 , RI1CLtat1aln et qua e h~ an d'oan n h'om mau Rh Thai Rh sau sinh (+) (+) (+) (+) (+) (+) (-) (-) 8 (+) (+) (+) (+) (+) (+) 8 Mil b~nh nhan RH08 RH09 RHIO RHII RH12 RHI3 -,j Tu6i thai Rh Thai Rh sau sinh , J ; 8 (+) (+) (+) (+) (+) l, (+) j (-) (-j i (-) (-) (+) (+) '1 i 1 l I Y HQC VI$T NAM THANG 11 • Tir k~t qua tren cho chung ta thay: J benh nhan nghien ciru c6 benh nhan mang thai mang nhom mau Rh( -) , cho thay khong co sir b~t d6nb nh6m mau gitra me va 'con (m~u Rh4, Rh 11 va Rh 12) C6 10 tong s6 13 m~u nghien ciru c6 thai nhi mang nh6m mau Rh(+), 10 m~u c6 sir b~t d6ng nh6m mau gitra me va Cac k~t qua da duoc xac djnh llili bang test nhanh nh6m mall Rh cua sau sinh Theo cac tac gia Aykut A,Onay H, Sagol S, Gunduz C,Ozkinay F, Cogulu et al (2013) nghien cuu chftn dean nh6m mall Rh cua thai nhi cho 29 tnrong hop me mang nh6m mau Rh () thAy xUc1thien 21 tnrong hqp nh6m mall Rh( +) va truang hgp mang nh6m mau Rh (-) Theo HlC gia Hahn S van der Schoot CE, Chitty LS (2008) dB eong chAn doan nhom mall Rh Clla thai nhi a cae ph\! nir mang thai co nhom mall Rh( -} [ Akolekar Finning, Kl1ppl1samy va e¢ng s\r f (20II) ch~n dOlin nhom mau Rh thai nhi cho ba m~ mang nhom ImilJ Rh( -) co thai tu tI cac tll~n thu 11 d~n tu6i thai 13 tu~n eho 502 truang hqp 496 trubng hgp ch§n doan chinh xae nhom mau eua thai nhi vai d¢ nh~y la 98.8% 338 tnrang tra lai duong t1nh thi co truang hqp la duang tinh gia Bombard Akolekar, Farkas va c¢ng s\r (2011) da nghien elm eh§n doan nhom mall Rh Clla thai nhi qlla mall m~ tren 207 tnrcmg hqp k~t qua 140 truemg hqp duang t1nh mang nhom mall Rh( +) nhlmg co tnrcmg hqp duong Hnh gia va 67 truang hgp am tinh d6 co tnrbng hqp am tinh gia I ! '" A IV KETLU~N Trong nghien elm chtmg toi dll sang 19C dugc 11 truang hqp gia dinh mang gen b~nh tang san thuqng th~n b§m sinh, 10 gia : dinh mang gen Teo co Duchenne va chfm 56 D~C 81$T/2014 doan duoc nh6m mau ella thai nhi a 13 phu nCrmang nh6m mall Rh (-) dang mang thai tuoi thai tuan thir va thir a TAl L1~UTHAM KHAo Akolekar, Kirstin Filming Ramesh Kuppusamy et al (2011) Fetal RHD Genotyping in Maternal Plasma at 11-13 Weeks of Gestation Fetal Diagn Ther ;29:301-306 Aykut A,Onay H, Sagol S, Guncluz C,Ozldnay F, Cogulu et al (2013) d status by Determination of fetal rhesus maternal plasma DNA analysis Balkan f A4ed(iellel.16(2):33-8 Bombard, Akolel,ar, Fnrkus et HI (2011) relal RHD gl:notypc detection Ii'om circulating ccii-free fetal DNA in maternal plasma in non-sensitized RhO negative women frenal Diagl1 31: 802-808 Du ••J, ZOli X, Pan Y ct al (2009) Noninvasive prenatal diagnosis of congenital adrenal hyperplasia in cell -free fetal DNA with cold-peR de ES Ramos, P·395 Hahn S, van der Schoot CE, Chitty LS (2008) Non-invasive prenatal diagnosis and determination of fetal Rh statlls Semin Fetal Neollala/ Med 13:63-8 Seldzawa A., T Kimura, M Sasaki, S Nakamura, R Kob~lyashi, and T Sato (1996) Prenatal diagnosis of Duchenne muscular dystrophy using a single fetal nucleated erythrocyte in maternal blood Neurology 46: 1350-1353 Walknowslm, J., F A Conte, and M M Grumbach (1969) Practical and theoretical impl ications of fetal-maternal lymphocyte transfer Lancel, I: I II 'i-I 122 1.37 ... thong qua ADN ph6i thai W mall ngo~i vi Clla my d6i v6i 13 tnrang hqp chAn doan va duQ'c chAn doan 13S CHUYEN DE: HiNH THAI H9C kiem tra chinh xac lai sau sinh Viec chan doan Rh sau sinh diroc thuc... Clla thai kY· II DOl TU'Q'NG, HOA CHAT VA PHU'ONG PHAp NGHIEN C(rU 2.1 DBi tU'C~)'Ognghien cU'u d nghien Clru Clla chung t6i, chlmg toi Iva ch9n thai di~ m cac tu6i thai til' tll~n thu cua thai. .. thU'Qngth~n bim sinh Sau da chuan hoa diroc ky thuat tach ::hi~t ADN phoi thai W va k5' thuat PCR nhfm gen SR Y chung toi tien hanh thir nghi~m sang loc benh tang san thuong th~n bam sinh tnroc sinh tuftn