Isolated Nonsyndromic Congenital Heart Disease: NKX2-5 Gene Sequencing

Condition Description

Mutations in the NKX2-5 gene have been associated with atrioventricular (AV) conduction block, septal defects, conotruncal abnormalities (particularly Tetrology of Fallot), and AV valve formation defects. Mutations in NKX2-5 have been observed in autosomal dominant pedigrees and isolated cases of congenital heart disease. Studies suggest that NKX2-5 mutation may be a frequent cause (up to 4%) of sporadic and familial congenital heart defects.

NKX2-5 mutation analysis is appropriate for patients with an atrioventricular conduction block or structural heart defects with or without a family history of congenital heart defects. Analysis includes sequencing the entire NKX2-5 coding region (2 exons) and immediate exon/intron boundaries. Mutations in other regulatory regions and large deletions will not be detected by this assay. Variants of unknown clinical significance may be detected. Custom mutation detection is available for known familial mutations. For patients with mutations not identified by full gene sequencing, a separate deletion/duplication assay is available using a targeted CGH array KP.

Genes (1)

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This test is indicated for:
  • Confirmation of a clinical diagnosis of NKX2-5-related congenital heart disease.
  • Carrier testing in adults with a family history of NKX2-5-related congenital heart disease.


The 2 exons and flanking regions of NKX2-5 are amplified by PCR and sequenced in both the forward and reverse directions. Custom mutation detection is available for known familial mutations.


Over 95% of mutations in the coding regions or splice sites will be detected by this assay. This assay will detect sequence variants in the coding region and splice junctions. Mutations in the promoter region, some mutations in the introns, and other regulatory elements cannot be detected by this analysis. Large deletion and insertion mutations will not be detected by this assay. It is possible that some patients with a typical presentation may not carry a mutation detected by this analysis. This analysis may detect novel variants of unclear effect, which may require further studies.

Specimen Requirements

Listed below are EGL's preferred sample criteria. For any questions, please call 470.378.2200 and ask to speak with a laboratory genetic counselor (
Submit only 1 of the following specimen types

Oragene™ Saliva Collection Kit
Orangene™ Saliva Collection Kit used according to manufacturer instructions. Please contact EGL for a Saliva Collection Kit for patients that cannot provide a blood sample.
Collection and Shipping
Please do not refrigerate or freeze saliva sample. Please store and ship at room temperature.
Whole Blood (EDTA)

EDTA (Purple Top)
Infants and Young Children (<2 years of age): 2-3 ml
Children > 2 years of age to 10 years old: 3-5 ml
Older Children & Adults: 5-10 ml
Autopsy: 2-3 ml unclotted cord or cardiac blood
Collection and Shipping
Ship sample at room temperature for receipt at EGL within 72 hours of collection. Do not freeze.
DNA, Isolated

Isolation using the Perkin Elmer™Chemagen™ Chemagen™ Automated Extraction method or Qiagen™ Puregene kit for DNA extraction is recommended.
Collection and Shipping
Refrigerate until time of shipment in 100 ng/µL in TE buffer. Ship sample at room temperature with overnight delivery.

Special Instructions

Submit copies of diagnostic biochemical test results with the sample. Sequence analysis is required before deletion/duplication analysis by targeted CGH array. If sequencing is performed outside EGL Genetics, please submit a copy of the sequencing report with the test requisition. Contact the laboratory if further information is needed.

How to Order

Requisition Forms