Hearing Loss: GJB2 & GJB6 Gene Sequencing Panel

Condition Description

Notice: This test has been discontinued.

NTD Genetics no longer accepts samples for this test. Please call: 470-378-2200 for assistance in identifying alternative testing options.

In the United States, approximately 1 in 1000 children are diagnosed with prelingual hearing loss (HL) or deafness. Approximately half of prelingual hearing loss or deafness is attributed to environmental exposures and the remaining half to genetic causes. Approximately 30% of hereditary hearing loss is estimated to be syndromic (associated with other birth defects) while the remaining 70% is non-syndromic (isolated and not associated with other findings). Non-syndromic deafness is mainly due to recessive genes (75-80%) and over 20 such genes have been identified, but non-syndromic deafness may also be inherited in autosomal dominant, X-linked, or mitochondrial patterns.

Molecular testing can aid in rapid diagnosis of hearing loss. Early diagnosis of hearing defects can provide diagnostic information, facilitate timely intervention, and assist with genetic counseling.

Connexins are transmembrane proteins that form channels that allow rapid transport of small molecules between cells; the proteins connexin 26 and connexin 30 interact to form a channel that functions in the inner ear. The GJB2 gene encodes the connexin 26 protein and is involved in 50% of autosomal recessive hearing loss. The GJB6 gene is located near GJB2, and encodes the protein connexin 30. Patients with non-syndromic hearing loss have been found to have two mutations in connexin 26, two mutations in connexin 30, or compound heterozygosity for one mutation in connexin 26 and another in connexin 30 [1,2].

This test panel includes complete sequencing of the genes for connexin 26 and connexin 30, and testing for the common 342kb deletion in connexin 30.

Please click here for the GeneReviews summary on this condition.


  • Pandya A, et al. (2003) Frequency and distribution of GJB2 (connexin 26) and GJB6 (connexin 30) mutations in a large North American repository of deaf probands. Genet Med 5:295-303.
  • Del Castillo I, et al. (2003). Prevalence and evolutionary origins of the del(GJB6-D13S1830) mutation in the DFNB1 locus in hearing-impaired subjects: a multicenter study. Am J Hum Genet 73:1452-8.

Genes (2)

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This test is indicated for:

  • Individuals with clinical findings consistent with non-syndromic hearing loss when mitochondrial etiologies have been ruled out.
  • For carrier testing of individuals with a family history of non-syndromic hearing loss.


PCR amplification of the exons and flanking regions contained in the GJB2 and GJB6 genes are performed on the patient's genomic DNA. Direct sequencing of amplification products is performed in both forward and reverse directions using automated fluorescence dideoxy sequencing methods. The patient's gene sequences are then compared to a normal reference sequence. Sequence variations are classified as mutations, benign variants unrelated to disease, or variations of unknown clinical significance. Variants of unknown clinical significance may require further studies of the patient and/or family members. This assay does not interrogate the promoter region, deep intronic regions, or other regulatory elements, and does not detect large deletions. The GJB6 gene 342kb deletion is detected by allele-specific amplification.


GJB2 Sequencing:
Detects over 98% of sequence variants in the coding region and splice junctions.

GJB6 Sequencing:
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. Mutations in the promoter region, some mutations in the introns, and other regulatory elements cannot be detected by sequence analysis. Large deletions such as the most common 342 kb deletion, and insertions will not be detected by sequence analysis.

GJB6 Deletion:
Will detect nearly all 342kb common deletion alleles in Connexin 30. Other deletion mutations reported in GJB6, including the 232kb (reported in Spain), 309kb (reported in UK), 140kb, and 150kb deletions will be detected by the separate GJB6 deletion/duplication array.

Specimen Requirements

Listed below are EGL's preferred sample criteria. For any questions, please call 470.378.2200 and ask to speak with client services (NTD_ATL_CS@eurofins-ntdgenetics.com).
Submit only 1 of the following specimen types
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.
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.

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.
  • The Hearing Loss: Comprehensive Panel (HL) is indicated for patients who have not have previous molecular testing and includes sequencing of the GJB2 and GJB6 genes, targeted mutation analysis of the GJB6 common 342kb deletion, and testing for mitochondrial mutations associated with aminoglycoside sensitivity.
  • For patients with mutations not identified by full gene sequencing, Hearing Loss: Connexin 26 and 30 Deletion/Duplication (PI) is available.
  • Known Mutation Testing (KM) is available to family members if mutations are identified by sequencing.
  • Mitochondrial-Related Hearing Loss: Mutation Panel (QJ) for patients with a history of aminoglycoside sensitivity.
  • Prenatal testing is available to couples who are confirmed carriers of mutations. Please contact the laboratory genetic counselor to discuss appropriate testing prior to collecting a prenatal specimen.

How to Order

Requisition Forms