Rapid Prenatal Diagnosis of Trisomy 21 by Real-time Quantitative Polymerase Chain Reaction with Amplification of Small Tandem Repeats and S100B in Chromosome 21

다국어 초록 (Multilingual Abstract)

Trisomy 21 (Down syndrome) is the most common congenital anomaly, and it occurs in one out of 700-1000 births. Current techniques such as amniocentesis and chorionic villi sampling (CVS) require lengthy laboratory culture procedures and high costs. This study was undertaken to establish a rapid prenatal diagnosis of trisomy 21 using real-time quantitative polymerase chain reaction (PCR) of fetal DNA from amniotic fluid. Real-time quantitative PCR was performed with DNA templates obtained from 14 normal blood samples, 10 normal amniotic fluid samples, 14 Down syndrome blood samples, and 7 Down syndrome amniotic fluid samples. Primers for D21S167 and S100B of chromosome 21 were used. Primers that direct the amplification of the 165-bp fragment of the insulin-like growth factor (IGF)-1 gene on chromosome 12 using a PCR primer were included to generate an internal standard for quantitation. The relative levels of D21S167 and S100B were 2.6 and 2.4 times higher in the blood of Down syndrome patients than those in the control group. The differences between these two groups were statistically significant (p-values were 0.0012 and 0.0016, respectively). The relative levels of D21S167 and S100B were 2.1 and 2.7 times higher in the amniotic fluid of Down syndrome fetuses than those in the control group. The difference between these two groups was statistically significant (p-values were 0.0379 and 0.0379, respectively). Prenatal diagnosis of trisomy 21 by real-time quantitative PCR using STR (small tandem repeats) amplification of D21S167 and S100B is a useful, accurate and rapid diagnostic method. Furthermore, it may also be useful for prenatal diagnosis with fetal DNA from maternal blood, and for preimplantation genetic diagnosis and prenatal counseling.

참고문헌 (Reference)

1 "TG repeat polymorphism at the D21S167 locus" 776-83,

2 "Successful diagnosis of fetal gender using conventional PCR analysis of maternal serum" 47 : 41-6, 2001

3 "Same day diagnosis of Down's syndrome and sex in single cells using multiplex fluorescent PCR" 51 : 164-7, 1998

4 "Regional assignment of human liver-type 6-phosphofructokinase to chromosome 21q22 3 by using somatic cell hybrids and a monoclonal anti-L antibody" 34-40,

5 "Rates of chromosome abnormalities at different maternal ages" 282-5, obstetgynecol1981

6 "Rapid trisomy diagnosis (21, 18 and 13) using fluorescent PCR and short tandem repeats: applications for prenatal diagnosis and preimplantation genetic diagnosis." 15 : 266-75, 1998

7 "Rapid prenatal diagnosis of trisomy 21 by polymerase chain reaction associated analysis of small tandem repeats and S100B in chromosome 21" 13 : 361-6, 1998

8 "Rapid detection of trisomy 21 by quantitative PCR" 567-70,

9 "Rapid detection of trisomy 21 by homologous gene quantitative PCR" 99 : 364-7, 1997

10 "Quantitative real-time polymerase chain reaction for the core facility using TaqMan and the Perkin-Elmer/Applied Biosystems Division 7700 sequence detector" 10 : 11-6, 1999

11 "Quantitative analysis of fetal DNA in maternal plasma and serum implications for noninvasive prenatal diagnosis" 62 : 768-75, 1998

12 "Presence of fetal DNA in maternal plasma and serum" 350 : 485-7, 1997

13 "Prenatal identification of fetal genetics traits" 357 : 310-1, 2001

14 "Prenatal genetic diagnosis in 3000 amniocentesis" Epstein CJ 157-63,

15 "Large amounts of cell-free fetal DNA are present in amniotic fluid" 1867-9

16 "Improved diagnosis of Duchenne/ Becker muscular dystrophy" 613-9, jclininvest1990

17 "High sensitivity of fetal DNA in plasma compared to serum and nucleated cells using unnested PCR in maternal blood" 15 : 102-7, 2000

18 "Fetal gender determination in early pregnancy through qualitative and quantitative analysis of fetal DNA in maternal serum" 110 : 75-9, 2002

19 "Experience of first 510 cases in Korean" 906-15, koreanjobstetgynecol1993

20 "Diagnosis of down syndrome and other aneuploidies using quantitative polymerase chain reaction and small tandem repeat polymorphisms" 43-50,

21 "Detection of fetal Rhesus D and sex using fetal DNA from maternal plasma by multiplex polymerase chain reaction" 107 : 766-9, 2000

22 "Clinical usefulness of fluorescence in situ hybridization in the diagnosis of genetic disease" 45 : 1016-25, 2002

23 "Chorionic villus sampling clinical experience of the initial 750 cases" 22 : 143-9, 1996