Limitations
For this patient, only genes that were associated with reported clinical
features were analyzed. Therefore, this study cannot exclude the
possibility of mutations in genes not related to the given phenotype.
Also, the reported variants were only seen in exome data. Therefore,
validation of detected variants by another molecular method, such as
PCR-Sanger sequencing, is recommended. It is of value to note that theTCF3 gene has a pseudogene (TCF3P1 ) with a high degree of
sequence similarity. Therefore, it is very important to validate this
variant by another molecular method before any clinical decision-making.
It should be kept in mind that there are still many limitations in using
NGS itself and also in the interpretation of the obtained data. Among
them is the possibility of pathologic variants present in the genome
that are not detected by the technique. Also, Large duplications and
deletions, balanced translocations, inversions, ploidy changes,
uniparental disomies, and methylation alterations cannot be detected by
this method.