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.