Introduction
Trisomy 20p manifesting as intellectual disability and speech delay is a
rare genetic disorder1,2. The cases of trisomy 20p
previously reported mostly focused on the patients after birth and there
is little information about prenatal screening of trisomy 20p
considering the lack of effective detection technology.
Since 2011, NIPT has been introduced into clinical practice for the
detection of fetal trisomy 21, 18, and 13 via massively parallel
sequencing of cell-free DNA (cfDNA) in maternal
plasma3. The fetal CNVs can also be detected due to
the statistical power of the methods published and bigger sequencing
depth4. Using two to eight million reads, Chen et al.
detected fetal CNVs bigger than 10 Mb5. The trisomy
20p can be screened theoretically considering the short arm of
chromosome 20 is bigger than 10 Mb and the mean sequencing reads of NIPT
is bigger than 3.5 million reads.
Here we firstly report a case of de novo trisomy 20p detected by NIPT in
a 27 years pregnant women, confirmed by prenatal diagnosis.
Clinical report
A 27-year-old pregnant woman (G1P0) with gestational age (GA) of 16
weeks 2 days accepted NIPT in reproductive medicine center of renmin
hospital, hubei university of medicine. The couple with no special face
and normal intelligence is not consanguineous and both healthy. The
fetal nuchal translucency (NT) thickness is 1.2 mm with GA of 13 weeks.
5 ml blood was collected in cfDNA storage tube (CW2814M) from the
pregnant woman. The cfDNA was extracted from plasma, followed by end
repair, adaptors ligation and PCR amplification according to the
instruction. Single-ended sequencing was performed on BGISEQ-500
(BGI-Wuhan, Wuhan, China) with the 35 bp length. The raw sequence reads
is 11.91 million, of which 74.2% is blasted to the human genome. The
effective reads is 7.98 million and the fetal fraction in the maternal
plasma is 9.35%. The Z-score of T21, T18 and T13 were less than 3 which
suggested 21-trisomy, 18-trisomy and 13-trisomy were negative while the
Z value of T20 is 5.508 which meaned that there was about a dupication
of chromosome 20 (Figure 1A).
Following genetic counselling, the amniocentesis was performed at 17
weeks of gestation under the guidance of ultrasound. STR analysis via
Multiple STR genotyping kit (DARUI biotechnology) suggested there was no
maternal cell contamination in amniotic fluid which was used to cell
culture analysis of fetal amniotic fluid karyotype and CNVs analysis by
LC-WGS6-8.
Cytogenetic analysis by G-banding shows the karyotype of the fetus with
46, XX, add(21)(p12) (Figure 1B). The karyotype analysis of the couple
showed no obvious abnormalities.
CNVs sequencing was performed on a BGISeq-500 platform (BGI-Wuhan,
Wuhan, China) as previously reported. In brief, 1μg genomic DNA
extracted from fetal amniotic fluid cell was randomly fragmented.
Library were conducted according to the standard instruction and
sequenced with a minimum of ~15 million reads
(single-end 50 bp) per sample8. The
homozygous/heterozygous duplication and deletion was detected by
reported method6. The CNV-Seq analysis results was
seq[GRCh37/hg19]20p11.22p13 (60,001-21,474,604)×3, indicating a 21.4
Mb duplication on chromosome 20p11.22-p13 (Figure 1C).
At 22 weeks of gestation, an isolated choroid plexus cyst measuring
0.41cm x 0.36cm in fetal brain was identified during routine sonography.
Discussion
Since the 1980s, chromosomal microdeletions/microduplications have been
be identified to associated with developmental delay and intellectual
disability9. The common use of next-generation
sequencing (NGS) and array comparative genomic hybridization (aCGH) in
diagnostics has led to an increase of known pathogenic CNVs10. Thanks to the application of NIPT, fetal
chromosomal aneuploidies subchromosomal abnormalities can be screening
with high sensitivity and specificity 11,12.
Trisomy 20p is a rare chromosomal duplication manifesting as poor
coordination, mental retardation with speech delay, and moderate facial
dysmorphisms which emerged from post-natal
observations13,14. There is only one prenatal case of
trisomy 20p resulting from maternal inversion
[46,XX,inv(20)(p11.2q13.3)] in chromosome 20. The only
ultrasonographic anomaly was an increased NT of 3.6 mm at 12
weeks15.
Here we report firstly a case of prenatal diagnosis of de novo trisomy
20p result from the parents with normal karyotype. The fetal karyotype
was 46, XX, add(21)(p12), indicating additional unknown origin was
attached to band 21p12. The CNV-Seq result revealed the additional
unknown origin may be 20p11.22-p13. The only ultrasonographic anomaly at
21 weeks was an choroid plexus cyst located on the left side of the
brain while the NT value is normal. The fetus after induction had normal
phenotypes. According to post-natal phenotypes described, prenatal
diagnosis of such partial trisomy 20p seems to be difficult, considering
the capacity of ultrasound to detect mild dysmorphy. However, NIPT has
good performance in detecting fetal trisomy 20p combined with LC-WGS of
amniotic fluid cell.
Further via deeper sequencing fetal subchromosomal CNVs can be detected
by NIPT for the lower sequencing cost. NIPT, combined with karyotype
analysis and NGS may be a efficient screening strategy for fetal trisomy
20p syndrome.
Acknowledgements: We thank the family for their participation
and for allowing us to report their family’s data.