Obstet Gynecol Sci.  2016 Nov;59(6):444-453. 10.5468/ogs.2016.59.6.444.

Quantitative fluorescent polymerase chain reaction for rapid prenatal diagnosis of fetal aneuploidies in chorionic villus sampling in a single institution

• Shin YJ
• Chung JH
• Kim DJ
• Ryu HM
• Kim MY
• Han JY
• Choi JS

• KMID: 2378568
• DOI: http://doi.org/10.5468/ogs.2016.59.6.444

Abstract

OBJECTIVE
To validate quantitative fluorescent polymerase chain reaction (QF-PCR) via chorionic villus sampling (CVS) for the diagnosis of fetal aneuploidies.
METHODS
We retrospectively reviewed the medical records of consecutive pregnant women who had undergone CVS at Cheil General Hospital between December 2009 and June 2014. Only cases with reported QF-PCR before long-term culture (LTC) for conventional cytogenetic analysis were included, and the results of these two methods were compared.
RESULTS
A total of 383 pregnant women underwent QF-PCR and LTC via CVS during the study period and 403 CVS specimens were collected. The indications of CVS were as follows: abnormal first-trimester ultrasonographic findings, including increased fetal nuchal translucency (85.1%), advanced maternal age (6.8%), previous history of fetal anomalies (4.2%), and positive dual test results for trisomy 21 (3.9%). The results of QF-PCR via CVS were as follows: 76 (18.9%) cases were identified as trisomy 21 (36 cases), 18 (33 cases), or 13 (seven cases), and 4 (1.0%) cases were suspected to be mosaicism. All results of common autosomal trisomies by QF-PCR were consistent with those of LTC and there were no false-positive findings. Four cases suspected as mosaicism in QF-PCR were confirmed as non-mosaic trisomies of trisomy 21 (one case) or trisomy 18 (three cases) in LTC.
CONCLUSION
QF-PCR via CVS has the advantage of rapid prenatal screening at an earlier stage of pregnancy for common chromosomal trisomies and thus can reduce the anxiety of parents. In particular, it can be helpful for pregnant women with increased fetal nuchal translucency or abnormal first-trimester ultrasonographic findings.

Keyword

Chorionic villi sampling; Fluorescence; Polymerase chain reaction; Prenatal diagnosis

MeSH Terms

Aneuploidy*
Anxiety
Chorion*
Chorionic Villi Sampling*
Chorionic Villi*
Cytogenetic Analysis
Diagnosis
Down Syndrome
Female
Fluorescence
Hospitals, General
Humans
Maternal Age
Medical Records
Mosaicism
Nuchal Translucency Measurement
Parents
Polymerase Chain Reaction*
Pregnancy
Pregnant Women
Prenatal Diagnosis*
Retrospective Studies
Trisomy

Figure

• 

  Fig. 1 Normal quantitative fluorescent polymerase chain reaction results with peak height ratios of allele dosage between 0.8 and 1.4 in more than two short tandem repeats (arrow head) in 46, XY, based on at least two informative markers, except for two short tandem repeat markers on chromosome 18 (D18S391, D18S390) showed uninformative single peaks (arrow). This case was confirmed as 46, XY, normal fetus by long term culture. The XY male sex chromosome constitution is identified by the presence of both the X- and Y- specific products of AMXY (with a normal ratio of 1:1) in addition to the SRY product and the single X-linked HPRT allele. AMXY, amylogenin XY; X, X chromosome; HPRT, hypoxanthine-guanine phosphoribosyltransferase; D, DNA; S, Segment; SRY, sex-determining region Y; Y, Y chromosome; ar,area; ht, height; sz, size.

• 

  Fig. 2 Positive quantitative fluorescent polymerase chain reaction results with the main short tandem repeat markers in 47, XY, +21: Trisomy 21 is determined by the trisomic triallelic pattern for D21S1414, D21S1411, D21S1435, and D21S1446 (arrow). AMXY, amylogenin XY; X, X chromosome; HPRT, hypoxanthine-guanine phosphoribosyltransferase; D, DNA; S, Segment; SRY, sex-determining region Y; Y, Y chromosome; ar,area; ht, height; sz, size.

• 

  Fig. 3 Positive quantitative fluorescent polymerase chain reaction results with extra short tandem repeat markers in 47,XY,+21: Trisomy 21 is identified by a trisomic diallelic pattern for D21S1008, D21S1412, and D21S1437 (≤0.6 or ≥1.8) (arrow) and a trisomic triallelic pattern for D21S1411 and D21S1435 (ratio 1:1:1) (arrow head). D, DNA; S, Segment; ar,area; ht, height; sz, size.

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