유전성 대사질환의 착상전 유전진단

강인수,Kang, Inn Soo 대한유전성대사질환학회 2005 대한유전성대사질환학회지 Vol.5 No.1

Prenatal diagnosis (PND) such as amniocentesis or chorionic villi sampling has been widely used in order to prevent the birth of babies with defects especially in families with single gene disorderor chromosomal abnormalities. Preimplantation genetic diagnosis (PGD) has already become an alternative to traditional PND. Indications for PGD have expanded beyond those practices in PND (chromosomal abnormalities, single gene defects), such as late-onset diseases with genetic predisposition, and HLA typing for stem cell transplantation to affected sibling. After in vitro fertilization, the biopsied blastomere from the embryo is analyzed for single gene defect or chromosomal abnormality. The unaffected embryos are selected for transfer to the uterine cavity. Therefore, PGD has an advantage over PND as it can avoid the risk of pregnancy termination. In this review, PGD will be introduced and application of PGD in inborn error metabolic disorder will be discussed.

골반결핵으로 인한 불임 환자에서 체외수정 시술의 결과

김진영,강인수,Kim, Jin-Yeong,Kang, Inn-Soo 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.4

Objectives: Pelvic tuberculosis (TB) causes infertility despite of anti-TB chemotherapy and IVF-ET is effective treatment to achieve pregnancy. The aim of this study is to assess the outcomes of IVF-ET in pelvic TB, especially according to main Tb lesion, and to investigate the factors affecting the successful outcome. Methods: A total of 135 IVF-ET cycles were performed in 54 patients with pelvic TB and the outcome was compared with that of control group with tubal factor not associated with TB in 301 cycles, 227 patients. Anti-TB chemotherapy was performed in the patients with pelvic TB. Pregnancy rate was compared according to main TB lesion as salpingitis, peritonitis, and endometritis. In the patients with endometrial TB, when complicated with uterine synechia, hysteroscopic lysis was done before IVF-ET and pregnancy rate was compared according to the presence of uterine synechia. Results: There was no significant difference in peak E2 ($2,790{\pm}280.1$ vs $2,554{\pm}101.2$, p>0.05), the number of retrieved oocytes ($13.5{\pm}0.7$ vs $12.5{\pm}0.4$, p>0.05) and fertilized oocytes ($7.7{\pm}0.5$ vs $7.8{\pm}0.3$, p>0.05) between patient and control group. Clinical pregnancy rate per transfer in pelvic TB group was 22.9% and showed no difference from that of control group (24.3%, p>0.05). Although it was not statistically significant, pregnancy rate in the endometrial Tb (18%) was lower than that in the salpingitis (28.5%) or peritonitis (26.5%) (p>0.05). In the endometrial TB with uterine synechia, pregnancy rate was significantly lower than that of the patients without synechia even after hysteroscopic lysis (9.7% vs 31.6%, p<0.05). Conclusion: IVF-ET after anti-TB chemotherapy is the most effective treatment to achieve pregnancy in infertile patients with pelvic TB. Because the presence of endometrial TB and resulting uterine synechia affects the outcome of IVF-ET, thorough evaluation for endometrium with endometrial biopsy and hysteroscopy is important to predict the prognosis of IVF-ET treatment.

자궁내막증이 있는 불임 여성에서 중등도에 따른 체외 수정의 결과 비교

김혜옥,강인수,Kim, Hye Ok,Kang, Inn Soo 대한생식의학회 2006 Clinical and Experimental Reproductive Medicine Vol.33 No.4

목 적: 자궁내막증이 체외 수정에 미치는 영향에 대하여 알아보고, 자궁내막증의 체외 수정 결과의 차이에 대해 살펴보고자 하였다. 연구방법: 1994년부터 2004년까지 제일병원 아이소망센타에서 자궁내막증으로 체외 수정을 시술 받은 697명의 환자 (총 1,199주기)를 후향적으로 연구하였다. 경증의 자궁내막증은 638주기, 중증의 자궁내막증은 561주기였으며, 난관 요인을 가진 325명 (459주기)를 대조군으로 하였다. 제외 기준으로는 여성의 나이가 35세 이상, basal FSH level이 20 mIU/ml 이상인 경우, 심각한 남성 요인의 경우를 제외하였다. 결 과: 중증의 자궁내막증은 난관 요인 보다 획득된 난자의 수 ($9.97{\pm}7.2$ vs. $13.4{\pm}7.9$ (p<0.0001)), 총 배아 수 ($6.5{\pm}4.8$ vs. $9.1{\pm}5.6$ (p<0.0001)), 양질의 배아 수 ($2.43{\pm}1.6$ vs. $2.74{\pm}1.7$ (p=0.013))가 통계적으로 유의하게 낮았다. 하지만, 중증의 자궁내막증의 임신율은 난관 요인과 유사하였다 (35.7 vs. 36.8 (%)). 경증의 자궁내막증은 중증의 자궁내막증과 난관 요인보다 불임 기간이 길었으며 ($55.4{\pm}25.7$ vs. $47.6{\pm}25.6$ vs. $44.4{\pm}30.9$ (개월) p<0.0001)), 수정률이 의미 있게 낮았으나 ($64.8{\pm}22.9$ vs. $69.9{\pm}22.5$ vs.$70.8{\pm}20.8$ (%) (p<0.0001)), 임신율에 유의한 차이는 없었다 (31.1 vs. 35.7 vs. 36.8 (%)). 또한, 체외 수성 이전에 치료 받은 병력이 있던 경증과 중증의 자궁내막증 (363주기 vs. 470주기)은 경증의 자궁내막증에서 중증의 자궁내막증보다 불임 기간이 길고 ($56.5{\pm}26.3$ vs. $46.9{\pm}25.8$ (개월), p<0.0001), 수정률이 낮았으며 ($64.7{\pm}23.3$ vs. $70.5{\pm}22.7$ (%), p=0.001), 임신율과 태아 생존율이 통계적으로 유의하게 낮았다 (29.2 vs. 36.2 (%), p=0.045, 23.9 vs. 31.5 (%), p=0.043). 결 론: 체외 수정 시 이전에 치료를 받았던 경증의 자궁내막증은 중증의 자궁내막증보다 임신율과 태아 생존율이 낮았고, 이는 현저한 수정률 감소와 긴 불임 기간이 관련이 있다고 생각된다. 따라서, 장기간의 불임 기간을 가진 경증의 자궁내막증을 가진 불임 여성은 체외 수정을 좀 더 일찍 고려해 볼 수 있겠다. Objective: To evaluate the impact of endometriosis on IVF-ET cycles and to compare IVF outcomes between stage I/II and stage III/IV endometriosis. Methods: We analyzed 697 patients (1,199 cycles) with endometriosis (stage I-II:638 cycles, stage III-IV: 561 cycles) and 325 pts (459 cycles) with tubal factor as controls between January 1994 and April 2004. Pts with endometriosis were diagnosed by laparoscopy and medical and surgical treatment were done in 353 cycles (55.3%) and 466 cycles (83.1%) of stage I-ll/stage III-IV endometriosis. Cycles with age>35 years or FSH>20 miU/mL or severe male factor infertility were excluded. Results: The number of retrieved oocytes ($9.97{\pm}7.2$ vs. $13.4{\pm}7.9$ (p<0.0001 )), total number of embryos ($6.5{\pm}4.8$ vs. $9.1{\pm}5.6$ (p<0.0001)), and good quality embryos ($2.43{\pm}1.6$ vs. $2.74{\pm}1.7$ (p=0.013)) significantly decreased in stage III-IV endometriosis than in control. But pregnancy rate of stage III-IV endometriosis was comparable with control (35.7% vs. 36.8%). Fertilization rate and number of total embryos were lower in stage I-II endometriosis than in control ($64.8{\pm}22.9$ vs. $70.8{\pm}20.8$ (p<0.0001), $7.6{\pm}5.0$ vs. $9.1{\pm}5.6$ (p<0.0001)). In patients with medical and surgical treatment of endometriosis, pregnancy rate and live birth rate was significantly lower in stage I-II than in stage III-IV endometriosis (29.2 vs. 36.2 (%), p=0.045, 23.9 vs. 31.5 (%), p=0.043). There was no difference in the mean age, but the duration of infertility was significantly longer ($56.5{\pm}26.3$ vs. $46.9{\pm}25.8$ (mon), p<0.0001) and fertilization rate was lower ($64.7{\pm}23.3$ vs. $70.5{\pm}22.7$ (%), p=0.001) in stage I-II than stage III-IV endometriosis. Conclusion: We suggest that IVF should be considered earlier in patients with minimal to mild endometriosis because of significantly decreased fertilization rates.

착상전 유전진단 (Preimplantation Genetic Diagnosis)의 임상적 적용

김진영,강인수,Kim, Jin-Yeong,Kang, Inn-Soo 대한생식의학회 2008 Clinical and Experimental Reproductive Medicine Vol.35 No.1

난소과자극증후군의 예측과 예방

김혜옥,강인수,Kim, Hye-Ok,Kang, Inn-Soo 대한생식의학회 2010 Clinical and Experimental Reproductive Medicine Vol.37 No.4

난소과자극증후군은 발생 시 생명을 위협하는 심각한 의인성 합병증으로, 불임 치료를 목적으로 성선자극호르몬을 사용하여 과배란을 유도할 때 발생한다. 따라서, 과배란 유도를 하기에 앞서 위험요인을 가진 환자를 파악하여 저용량의 성선자극호르몬을 사용하거나, GnRH antagonist protocol을 이용함으로써 발생을 예방하는 것이 중요하고, 과배란 유도 중 ovarian hyperstimulation syndrome (OHSS)의 발생이 예측될 때는 성선자극호르몬 투여 시 coasting을 하고 난포 성숙을 유도할 때 저용량의 hCG 혹은 GnRH agonist를 이용하고, 중증의 OHSS가 예측될 때에는 주기취소로 OHSS의 유병기간을 줄이거나 배아동결 등을 통해 후발성 난소과자극증후군을 예방할 수 있다. 그리고, metformin과 dopamine agonist를 난소과자극증후군을 예방을 위해 고려해 볼 수 있겠다. Ovarian hyperstimulation syndrome (OHSS) is a life-threatening iatrogenic complication of ovulation induction. Before ovarian stimulation, identification of patients vulnerable to developing OHSS is necessary. And ovarian stimulation should be started with low doses of gonadotropin or GnRH antagonist protocol. During monitoring of ovarian stimulation with risk of OHSS, coasting, low doses hCG and GnRH agonist for triggering ovulation are considered. If severe OHSS is predicted, cycle cancellation and cryopreservation of all embryos should be considered to reduce late-onset OHSS and morbidity. And metformin and dopamine agonist for reducing OHSS are being proposed as a prophylactic treatment for OHSS.

비정상 산과력을 가진 부부에서의 균형전좌형 염색체 보인자의 빈도 및 그 보인자들에서의 산전 세포유전학적 진단

박소연,강인수,류현미,전종영,이문희,김진미,최수경,Part, So-Yeon,Kang, Inn-Soo,Ryu, Hyun-Mee,Jun, Jong-Young,Lee, Moon-Hee,Kim, Jin-Mi,Choi, Soo-Kyung 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.3

Cytogenetic analysis was performed in 1321 couples and 141 women with history of abnormal reproductive outcome during 1988-1996. The use of high resolution banding technique and fluorescence in situ hybridization (FISH) in the chromosome analysis has made the precise evaluation of chromosome aberrations. The prevalence of balanced chromosomal translocation carriers were 3.74% (104/2783 patients). 70 cases (2.52%) were reciprocal translocation carriers and 34 (1.22%) had Robertsonian translocations. Chromosome aberrations were more frequent in women (73 cases) than in men (31 cases). No phenotypical abnormalities were found in all carriers, but they experienced abnormal reproductive outcomes such as recurrent spontaneous abortions, anomalous offsprings or infertility problem. Prenatal diagnosis was carried out on 36 subsequent pregnancies in balanced translocation carriers. The fetal karyotypes showed that 12 cases (33%) were normal, 22 (61%) were balanced translocations, and two (6%) were unbalanced translocations. It is concluded that the prevalence of balanced chromosomal translocations in patients with abnormal reproductive outcome is higher than that of the normal population. Most of the fetal samples showed normal karyotypes or balanced translocations. Although the incidence of chromosomal imbalance in the fetuses was relatively low in prenatal diagnosis, individuals with balanced translocations are predisposed to abnormal offspring with partial trisomy or monosomy. Therefore we recommend that genetic counselling and cytogenetic prenatal diagnosis for translocation carriers have to be offered to prevent recurrent chromosomal abnormal babies.

남성 불임 환자에서 Y 염색체 미세 결손이 체외 수정 결과에 미치는 영향

최노미,양광문,강인수,서주태,송인옥,박찬우,이형송,이현주,안가영,한호섭,이희정,김나영,유승연,Choi, Noh-Mi,Yang, Kwang-Moon,Kang, Inn-Soo,Seo, Ju-Tae,Song, In-Ok,Park, Chan-Woo,Lee, Hyoung-Song,Lee, Hyun-Joo,Ahn, Ka-Young,Hahn, Ho-Suap,Lee, H 대한생식의학회 2007 Clinical and Experimental Reproductive Medicine Vol.34 No.1

목 적: 무정자증이나 심한 희소정자증이 있는 남성 불임 환자에서 Y 염색체 미세 결손을 동반하는 경우 체외 수정 및 세포질내 정자 주입술과 배아 이식의 결과에 미치는 영향을 알아보고자 하였다. 연구방법: 후향적 실험군 -대조군 연구로, Y 염색체 미세 결손이 동반된 무정자증이나 심한 희소정자증이 있는 남성 불임 부부 14 쌍을 실험군으로 하였고 대조군으로는 남성 배우자가 무정자증이나 심한 희소정자증이 있지만 Y 염색체는 정상인 12 쌍의 불임 부부를 대상으로 하였으며, 이 두 군을 다시 정자 채취 방법에 따라 사정된 정자를 이용한 군과 고환 내 정자 추출술을 통해 정자를 채취한 군으로 분류하였다. 연구 결과로는 수정률, 등급이 우수한 배아 수, 착상률, 융모막성 성선자극호르몬 양성률, 초기 임신 소실률, 출생률을 비교하였다. 결 과: 등급이 우수한 배아 수, 착상률, 융모막성 성선자극호르몬 양성률, 초기 임신 소실률, 출생률은 Y 염색체 미세 결손이 있는 군과 정상 Y 염색체를 갖는 군 간에 차이가 없었으나, 수정률은 Y 염색체 미세 결손이 있는 불임 부부 (61.1%) 에서 정상 Y 염색체를 갖는 불임 부부 (79.8%, p=0.003) 보다 낮았다. 정자 채취의 방법에 따라 비교하였을 때, 고환 내 정자 추출술을 통해 정자를 채취한 경우 실험군 (52.9%) 에서 대조군 (79.5%, p=0.008) 보다 유의하게 수정률이 낮게 나타났으나 사정된 정자를 이용한 경우에는 실험군에서 대조군 보다 수정률이 더 낮은 경향을 보였으나 유의한 차이는 없었고, 나머지 결과들은 두 군 간에 차이가 없었다. 결 론: Y 염색체 미세 결손이 동반된 무정자증이나 심한 희소정자증이 있는 불임 부부에서 고환 내 정자 추출술을 통해 정자를 채취할 경우 체외 수정 및 세포질내 정자 주입술 결과 정상 Y 염색체를 갖는 불임 부부에서 보다 수정률이 더 낮게 나타났으나, 사정된 정자를 이용하는 경우나 고환 내 정자 추출술을 통해 정자를 채취하여 수정이 된 후에는 Y 염색체의 미세 결손이 체외 수정 및 세포질내 정자 주입술과 배아 이식의 결과에 영향을 주지 않는 것으로 보인다. 그러나 향후 대규모의 전향적 연구가 더 필요할 것으로 사료된다. Objective: To determine whether the presence of Y-chromosome microdeletion affects the outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) program. Methods: Fourteen couples with microdeletion in azoospermic factor (AZF)c region who attempted IVF/ICSI or cryopreserved and thawed embryo transfer cycles were enrolled. All of the men showed severe oligoasthenoteratoazoospermia (OATS) or azoospermia. As a control, 12 couples with OATS or azoospermia and having normal Y-chromosome were included. Both groups were divided into two subgroups by sperm source used in ICSI such as those who underwent testicular sperm extraction (TESE) and those used ejaculate sperm. We retrospectively analyzed our database in respect to the IVF outcomes. The outcome measures were mean number of good quality embryos, fertilization rates, implantation rates, $\beta$-hCG positive rates, early pregnancy loss and live birth rates. Results: Mean number of good quality embryos, implantation rates, $\beta$-hCG positive rates, early pregnancy loss rates and live birth rates were not significantly different between Y-chromosome microdeletion and control groups. But, fertilization rates in the Y-chromosome microdeletion group (61.1%) was significantly lower than that of control group (79.8%, p=0.003). Also, the subgroup underwent TESE and having AZFc microdeletion showed significantly lower fertilization rates (52.9%) than the subgroup underwent TESE and having normal Y-chromosome (79.5%, p=0.008). Otherwise, in the subgroups used ejaculate sperm, fertilization rates were showed tendency toward lower in couples having Y-chromosome microdeletion than couples with normal Y-chromosome. (65.5% versus 79.9%, p=0.082). But, there was no significance statistically. Conclusions: In IVF/ICSI cycles using TESE sperm, presence of V-chromosome microdeletion may adversely affect to fertilization ability of injected sperm. But, in cases of ejaculate sperm available for ICSI, IVF outcome was not affected by presence of Y-chromosome AZFc microdeletion. However, more larger scaled prospective study was needed to support our results.

Efficacy of Coculture System in the Patients with Poor Prognoses on Human IVF-ET Program

변혜경,염혜원,궁미경,손일표,강인수,이호준,Byun, Hye-Kyung,Youm, Hye-Won,Koong, Mi-Kyung,Son, Il-Pyo,Kang, Inn-Soo,Lee, Ho-Joon The Korean Society for Reproductive Medicine 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.2

본 연구는 Vero cell을 이용한 사람 배아의 공동배양술이 배아의 질을 향상시킬 수 있거나 또는 반복적 착상 실패를 극복하여 임신을 가능케 할 수 있는지 알아 보고자 시행되었다. 1996년 일년 동안, 반복적 착상 실패를 경험한 환자 (group I)와 이전 주기에서 배아의 질이 나빴던 환자 (group II)를 포함한 총 202례를 분석하여 대조군과 공동배양군 간의 배아의 질, 임신률, 임신유지율 및 착상률을 비교하였다. Group I 93례 가운데 34례는 공동배양을, 나머지 59례는 기존의 체외수정을 시행하였다. Group II 109례에서는 공동배양 36례, 기존의 체외수정 73례를 시행하였다. Group I에서 공동배양군의 임신률, 임신유지율 및 착상률은 각각 14/34 (41.2%), 9/34 (26.5%), 16/81 (19.8%)로 대조군 (11/59 (18.6%), 8/59 (13.6%), 12/152 (7.9%))에 비하여 높았으며, 특히 임신률과 착상률은 유의한 차이를 나타내었다(p=0.028, p=0.015). Group II에서는 공동배양군의 임신률과 임신유지율 및 착상률이 각각 8/36 (22.2%), 5/36 (13.9%), 8/87 (9.2%)로 대조군 (5/73 (6.8%), 3/73 (4.1%), 3/158 (1.9%))에 비하여 높았고, group I의 결과에서와 마찬가지로 임신률과 착상률의 유의한 차이를 나타내었다(p=0.028, p=0.022). 이상에서 Vero cell을 이용한 공동배양술은 위의 두가지 주소를 가진 환자군에서 좋은 결과를 나타내었다. 또한 group II에서 3일-공동배양군의 임신률 역시 향상되어 (4/15 (26.7%)), 보조부화술을 겸비한 3일-공동배양이 이전 주기에서 배아의 질이 나빴던 환자군에 적용될 수 있음을 알 수 있었다. 결론적으로 Vero cell을 이용한 공동배양술은 반복적 착상 실패를 경험한 환자나 또는 이전 주기에서 배아의 질이 나빴던 환자에게 적용하여 임신률을 향상시킬 수 있을 것으로 사료된다. The present study was carried out to evaluate whether the coculture system of human embryos with Vero cells can improve the quality of embryo or overcome the repetitive implantation failures in order to obtain pregnancy. From January to December 1996, a total 202 cases which patients with the problems of repetitive implantation failures (group I) or those with the poor embryonic quality in their previous cycles (group II) was analysed. The quality of cocultured embryo, pregnancy, on-going and implantation rates between coculture and control groups were compared. Of 93 cases in group I, coculture was performed in 34 cases and conventional IVF for the rest. Of 109 cases in group II, 36 for coculture and 73 for conventional IVF. In group I, pregnancy, on-going and implantation rates in coculture group (14/34 (41.2%), 9/34 (26.5%), 16/81 (19.8%), respectively) were higher than those of control (11/59 (18.6%), 8/59 (13.6%), 12/152 (7.9%), respectively). There is significance in the pregnancy and implantation rates (p=0.028 and p=0.015). In group II, pregnancy, on-going and implantation rates in coculture group (8/36 (22.2%), 5/36 (13.9%), 8/87 (9.2%), respectively) were higher than those of control (5/73 (6.8%), 3/73 (4.1%), 3/158 (1.9%), respectively). Like the result of group I, there is significance in the pregnancy and implantation rates (p=0.028 and p=0.022). Coculture system with Vero cells works well in the groups of the two indications. Although the case of 3 day-coculture was small as 15 cases in group II, 3 day-coculture improved pregnancy rate (4/15 (26.7%)). Therefore, 3 day-coculture with assisted hatching is recommended to the patients with poor embryonic quality. In conclusion, coculture system with Vero cells can be suggested as an effective method which improves pregnancy rate in those who have repetitive implantation failures or whose embryonic quality was poor in their previous cycles.

Reliability of the Single Cell PCR analysis for Preimplantation Genetic Diagnosis of Single Gene Disorders

최혜원,이형송,임천규,궁미경,강인수,전진현,Choi, Hye Won,Lee, Hyoung-Song,Lim, Chun Kyu,Koong, Mi Kyoung,Kang, Inn Soo,Jun, Jin Hyun The Korean Society for Reproductive Medicine 2005 Clinical and Experimental Reproductive Medicine Vol.32 No.4

연구목적: 단일 유전자 이상에 대한 착상전 유전진단을 성공적으로 시행하기 위해서는 효과적이고 신뢰도가 높은 PCR 방법의 확립이 중요하다. 본 연구에서는 alkaline lysis와 duplex nested PCR 방법을 단일 림프구와 할구의 유전자 분석에 적용하여 그 효용성을 확인하고자 하였다. 재료 및 방법: 단일 유전자의 이상이 확인된 Duchenne muscular dystrophy (DMD), ornithine transcarbamylase (OTC) 결핍증과 epidermolysis bullosa (EB) 가계의 대상자들에서 채취한 단일 림프구와 공여 받은 배아의 할구를 이용하여 각각 PCR, restriction fragment length polymorphism (RFLP)와 direct DNA sequencing 분석을 시행하였다. 이러한 분석에서 유전자 증폭률 (amplification rate)과 두개의 allele 중에서 하나의 allele이 증폭되지 않는 allele drop-out (ADO) 빈도에 대해 살펴보았다. 결 과: 단일 림프구와 할구를 이용한 PCR 방법의 유전자 증폭률은 DMD에서 91.1%와 81.8%, OTC 결핍증에서 96.0%와 78.1%, EB에서 91.3%와 90.0%를 각각 나타냈으며, ADO 빈도는 OTC 결핍증에서 13.3%, EB에서 16.8%로 관찰되었다. 결 론: 본 연구에서 적용한 alkaline lysis와 duplex nested PCR 방법은 단일 유전자에 대한 착상전 유전진단에 성공적으로 적용할 수 있는 방법으로 생각되며, ADO 빈도를 최소화할 수 있는 효율적인 방법의 개발에 대한 지속적인 연구가 필요하다.

근이양증 가계에서의 PEP-PCR을 이용한 착상전 유전자진단

최수경,이은호,이호준,전진현,강인수,백은찬,류현미,전종영,Choi, Soo-Kyung,Lee, En-Ho,Lee, Ho-Joon,Jun, Jin-Hyun,Kang, Inn-Soo,Paik, Eun-Chan,Ryu, Hyun-Mee,Jun, Jong-Young 대한생식의학회 1996 Clinical and Experimental Reproductive Medicine Vol.23 No.1

General PCR technique alone has a limitation for preimplantation genetic diagnosis(PGD) using single blastomere. Recntly developed primer extension preamplification(PEP) technology amplifies the whole genome and thus, simultaneous multiple locus analysis became possible. In this study, we report the efficacy of PEP-PCR for PGD in three muscular dystrophy carriers undergoing IVF-ET. A total of 37 blastomeres were obtained from 40 embryos at six to eight cell stage in three IVF cycles in two DMD and one BMD carriers. Whole genome from single blastomeres were amplified using I5-base oligonucleotide random primers. PCR amplified products of exon 45 in the dystrophin gene and alphoid X/Y loci for gender determination were analysed by 2% metaphor gel electrophoresis. A total of 37 PEP-PCR replicates from 37 single blastomeres from 40 embryos and 37 blanks were performed. We obtained the reliable results for exon 45 and alphoid X/Y. Transfer of female embryos and unaffected male embryo was attempted in three couples. Unfortunately, pregnancy was not achieved in these cases. PEP-PCR is a reliable and efficient PGD method in multiple locus analysis using single blastomere.