Collect whole blood in a purple top (EDTA) tube.
Refrigerate sample until shipment. Send the sample at room temperature with overnight delivery for receipt Monday through Friday within 24 hours of collection.
Whole blood can be refrigerated until shipment.
Heparinized specimens, severely hemolyzed specimens, frozen, clotted or possibly commingled specimens, blood in non-sterile or leaky containers, mislabeled or inappropriately labeled specimens.
Do not heat, freeze or centrifuge blood before shipment. Refrigerate sample until shipment.
Mon - Fri 9:00am to 4:00pm
81401, 81404, 81405(x2), 81408, 81479(x12)
The craniofacial panel includes 19 genes that are known to be associated with craniosynostosis, frontonasal dysplasias, and skeletal dysplasias (see Molecular Genetics section for more details).
The craniosynostosis (CSS) syndromes are a group of disorders characterized by the premature fusion of sutures of the skull. Developmental delay, seizures and dysmorphisms involving the face, skeleton, and nervous system are commonly associated with craniosynostosis. Craniosynostosis occurs in one out of 2,200 live births and affects males twice as often as females. Genetic basis for craniosynostosis is heterogeneous and mutations in multiple genes involved in the FGFR signaling pathway are known to cause the disease.
This panel includes many disorders with a primary feature presentation being craniosynostosis, along with disorders that present with craniosynostosis as one of a larger collection of features (i.e. Marfan’s syndrome).
Frontonasal dysplasias are disorders characterized by hypertelorism, a flat, broad nose, median facial cleft, widow’s peak hairline, and lack of formation of the nasal tip. The prevalence of frontonasal dysplasia is not known with certainty, but it is a rare finding. Frontonasal dysplasia is heterogeneous and mutations in homeobox genes and ligands of receptor tyrosine kinases cause this phenotype.
Skeletal dysplasias are disorders characterized by shortening of bones in legs and/or arms, bowed or fractured bones, abnormal ribs, asymmetric bone growth, polydactyly, and in some skeletal dysplasias, craniosynostosis. The prevalence of skeletal dysplasia is one out of 4,000-5,000 live births. Cases including craniosynostosis are found to have mutations in genes involved in the FGFR signaling pathway.
The genes involved in the Craniofacial panel include TWIST1, FBN1, RECQL4, MSX2, FGFR1, FGFR2, FGFR3, POR, GLI3, EFNB1, RAB23, TGFBR1, TGFBR2, IFT122, ALX1, ALX3, ALX4, TCF12, and ERF. Mutations in these disorders can be inherited in an autosomal dominant, autosomal recessive, or X-linked pattern. (See Genes and Associated Syndromes Table)Genes and Associated Syndromes Table
Next Generation Sequencing for TWIST1, FBN1, RECQL4, MSX2, FGFR1, FGFR2, FGFR3, POR, GLI3, EFNB1, RAB23, TGFBR1, TGFBR2, IFT122, ALX1, ALX3, ALX4
Genomic DNA is extracted from blood or other patient tissues following standard DNA extraction protocols. This test is performed by next generation sequencing using the SureSelectXT Target Enrichment System (Agilent Inc) followed by Illumina MiSeq sequencing (2x 150bp paired end) of the coding regions and splice sites of the targeted genes. FASTQ data is de-multiplexed based on sample-specific index sequence using the MiSeq Reporter software and aligned to human reference genome hg19 using Novoalign. Variants are then called using the GATK pipeline and annotated using ANNOVAR and SnpEff.
Sanger sequencing is performed to confirm clinically significant variants and to fill in regions with insufficient coverage (bases with less than 30X coverage). Variants that are classified as benign or likely benign will not be confirmed.
Sanger sequencing for TCF12 and ERF
PCR amplification and sequencing is performed on all coding exons including splice junctions. The patient’s gene sequence is then compared to a reference sequence. Sequence variants are classified as mutations, variants of unknown significance or benign variants unrelated to disease. Variants of unknown significance may warrant further studies in the patient and other family members.
Variants of unknown significance may warrant further studies in the patient and other family members.
Test results with interpretation will be mailed and/or faxed to the referring physician or send out lab following completion of the test. Additional reports will be provided as requested.
The clinical utility of the assay is to support a clinical diagnosis of the disease, facilitate genetic counseling, and assess the risk to other first degree relatives and to facilitate testing of at - risk family members.
Whole blood in EDTA purple top tubes is the preferred sample. High molecular weight genomic DNA, cheek epithelial cells, or other samples containing DNA may be acceptable. Contact the laboratory for specific instructions regarding such samples before sending the sample.
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