Yonsei Med J.  2005 Apr;46(2):193-197. 10.3349/ymj.2005.46.2.193.

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

• Yang YH
• Nam MS
• Yang ES

• KMID: 1734043
• DOI: http://doi.org/10.3349/ymj.2005.46.2.193

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.

Keyword

Real-time quantitative PCR; trisomy 21; prenatal diagnosis

MeSH Terms

Amniotic Fluid/*chemistry
Biological Markers/analysis
Chromosomes, Human, Pair 21/*genetics
Computer Systems
Down Syndrome/*diagnosis
Female
Humans
Nerve Growth Factors/*analysis
*Polymerase Chain Reaction
Pregnancy
Prenatal Diagnosis/*methods
S100 Proteins/*analysis
*Tandem Repeat Sequences
Time Factors

Figure

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  Fig. 1 Relative DNA levels of D21S167 and S100B determined by real-time quantitative PCR in the blood of Down syndrome and normal control groups. The relative DNA levels of D21S167/IGF-1 in blood were 2.3 ± 0.4 in the normal control group (n=14) and 6.1 ± 0.8 in the Down syndrome patients group (n=14), and the relative DNA levels of S100B/IGF-1 were 6.4 ± 1.0 in the normal control group (n=14) and 15.6 ± 2.1 in the Down syndrome group (n=14, mean±SEM). Brackets indicate SEM. *p < 0.01. (Nor, Normal; DS, Down syndrome).

• 

  Fig. 2 Relative DNA levels of D21S167 and S100B determined by real-time quantitative PCR in the amniotic fluid of the Down syndrome and normal control groups. The relative DNA levels of D21S167/IGF-1 in amniotic fluid were 3.0 ± 1.1 in the normal control group (n=10) and 7.0 ± 2.5 in the Down syndrome patients group (n=7), and the relative DNA levels of S100B/IGF-1 were 6.3 ± 2.5 in the normal control group (n=10) and 19.1 ± 7.0 in the Down syndrome group (n=7, mean±SEM). Brackets indicate SEM. *p < 0.05. (Nor, Normal; DS, Down syndrome).

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