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Publicación Familia Trisomía 9p
Publicación Familia Trisomía 9p
INFORME CLINICO
II
II
I
IV
Afectado
s
Portadores de la translocación balanceada ( con estudio
citogenético)
Retraso Mental (no
explorados)
Patient 2 (Fig. ) Is the 9 years old brother with a similar clinical picture of patient 1.
He was born by cesarean section for fetal distress at 38 weeks gestation, his birth weight
was 2 800g, no other perinatal impairments were detected. He showed early
psychomotor developmental delay. At 24 months old he underwent a surgical correction
of an umbilical hernia. At 4 years old, after cryptorchidism diagnosis was made, left
orchidopexia and right orchiectomy were practiced him. At present evaluation, we
found him with the follow clinical abnormalities: weight 25K ( ), height 1.26m ( ),
cephalic circumference 48cm ( ). Microcephaly, frontal estrecho, round facies,
downslating palpebral fissures, ocular hiperthelorim, telecanthus and epicanthic folds,
broad and prominent nasal bridge, fleshy tip, short filtrum, downturned corners of the
month, narrow high palatal arch, micrognathia, Low set misshaped prominent ears,
short neck. Normal thorax witthout abnormal heart murmurs. Small hands,
brachymesodactyly of the fingers, fifth finger clinodactyly, hipoplastic nails, and
abnormal palmar creases. External genitals with hypoplastic scrotum and diminished
size of the left testicle.
Cont. Table
Strabism - -
Short Filtrum + +
Hypertelorism +
Microcephaly + +
Brachycephaly + +
Deep-set eyes - +
Narrow Palatal Arch + +
Downslating Palpebral Fisures + +
Micrognatia + +
Skelethal Anomalies + +
Short Stature + -
Diminished Skelethal Maduration - -
Fifth Finger Clinodactyly + +
Brachymesodactyly + +
Hypoplastic Phalanges + +
Hypoplastic Nails + +
Lordosis - -
Scholiosis - -
Short Neck + +
Dislocation of the hip + -
Congenital Heart Defect In course Not
explored
Ventricular Septal Defect
Ebstein Anomalie
Patent Ductus Arteriosus
Renal Malformations - -
Umbilical hernia (poco frecuente) - +
Materials and Methods
Cytogenetic analysis.
Cytogenetic analysis with a resolution of 350-500 bands from lymphocytes of samples
from the two siblings, the older brother, the parents, and the maternal aunt and uncle
were performed according to standard techniques. Chromosomes were analyzed with
routine karyotype, GTG-banding, NOR banding, CGB and FISH. For the FISH
analysis the single copy LSI TUPLE 1 and ARSA (VYSIS) probes were used as a
control for regions 22q11.2 and 22q13.
Microarray analysis
Peripheral blood samples were collected and DNA was extracted using the Qiagen Maxi
Kit according to manufacturers instructions (Qiagen Inc., Hidelberg, GE).
The concentration of genomic DNA for all samples was measured using a
spectrophotometer (NanoDrop ND-1000, NanoDrop Technologies). Genomic DNA
integrity was evaluated with a 1% agarose gel electrophoresis. Samples were adjusted to
a 50 g/l concentration for further microarray processing .
Microarray analysis was performed using the Affymetrix Genome-Wide Human SNP
Array 6.0 that includes a total of 906,600 single nucleotide polymorphisms (SNPs) and
more than 946,000 copy number variation probes. Array processing and hybridization to
was performed using the Genechip reagents kit and manufacturers instructions
(Affymetrix, Santa Clara, CA). Briefly, 250 ng of genomic DNA was digested with Sty
I and Nsp I (New England BioLabs) and then ligated to Sty and Nsp specific adaptors.
A generic primer that recognizes the adaptor sequence was used to amplify the DNA
fragments. DNA fragments are PCR amplified in a GeneAmp PCR System 9700
(Applied Biosystems). PCR products from both reactions were pooled and purified with
magnetic beads (Agencourt Magnetic Beads, Beckman). Concentration was measured
with a Multiskan Spectrum (Thermo,Inc) and only samples with a concentration over
3.5 g/l were further fragmented and labeled with biotin. Hybridization was done for
16 hrs at 50℃in the Affymetrix GeneChip Hybridization Oven 640. Microarrays were
washed and stained with a streptavidin-phycoeritrin solution (Molecular Probes,
Invitrogen) using the Genechip Fluidics Stations 450 (Affymetrix). The Genechip arrays
were scanned using the Affymetrix 3000 7G scanner and GeneChip Operating Software
version 1.4 was used to generate the .CEL intensity files. Quality control of all
microarrays were evaluated with the Genotyping Console version 3.0.2.
Analysis of CNVs
Genotyping Console 3.0.2 was used to assess quality control of the microarray
experiments. Results for Call rate and Contrast QC were the main parameters
considered. Genotyping was performed using the Birdseed v2 algorithm.
Allele copy number variation and segmentation was calculated with the Partek
Genomics Suite software, version 6.4 (Copyright© 2008, Partek Inc., St. Louis, MO,
USA). Copy number and allele differences were obtained by an unpaired analysis ,
comparing the test samples with a baseline generated from 75 healthy mexicans samples
genotyped with the same microarray platform. Allele Intensity import for baseline and
problem samples was done using the same parameters, so fragment length correction
was performed by Partek algorithm to avoid bias. The Partek genomic segmentation
algorithm detects segments in neighboring regions where there is statistically significant
(p value treshold <0,001) difference in average intensity. A minimum amount of 10
markers were considered for segmentation results.
All the metaphases studied in the proband and in her brother (proband 2), showed a
partial trisomy in chromosome 9 from 9pter to 9q13. GTC karyotype result was 46,XX
+der(9)t(9;22)(q13;q11)mat,-22. FISH analysis results were 46,XX,ish22q11.2
(D22S533,D22S942x2), 22q13.3 (ARSAx2) (ISCN,2005). The sample from the father
and older brother showed a normal karyotype. The karyotype of the mother showed that
she is a carrier of a balanced translocation (9;22)(q13;q11) (Figure 1). In the clinical
interview, where the mother stated the there was a possibility of finding other carriers of
the translocation in the family, a cytogenetic analysis was performed to samples from
the sister and brother (maternal line aunt and uncle) and they are also carriers of the
traslocation. A detailed interview to the members of the family was performed and it
was inferred that the grandmother and great-grandfather were also obligatory carriers of
the translocation.
Microarray results
The 1.02 Mb deletion of 5q13.2 is one of the interesting aberrations detected in both
affected children. (REVISAR GENES DE LA REGION)This telomeric region is part of
a 500Kb inverted duplication, containing four genes (SERF1A,SERF1B,SMN1,SMN2)
and repetitive elements, which make it prone to rearrangements and deletions. Smn1
and Smn2 genes code for the survival motor neuron protein (SMN), a molecule with
Many of these alterations are located in duplicated regions, prone to rearrangements and
other are known polymorphic variants such as the olfactory receptor genes (OR4S2,
OR11H12, OR11H1)