CHARGE syndrome is an autosomal dominant multisystem disorder caused by mutation 
in the CHD7 gene, encoding chromodomain helicase DNA-binding protein 7. 
Molecular diagnostic testing for CHD7 mutation has been available in a clinical 
setting since 2005. We report here the results from the first 642 unrelated 
proband samples submitted for testing. Thirty-two percent (n = 203) of patient 
samples had a heterozygous pathogenic variant identified. The lower mutation 
rate than that published for well-characterized clinical samples is likely due 
to referral bias, as samples submitted for clinical testing may be for 
"rule-out" diagnoses, rather than solely to confirm clinical suspicion. We 
identified 159 unique pathogenic mutations, and of these, 134 mutations were 
each seen in a single individual and 25 mutations were found in two to five 
individuals (n =69). Of the 203 mutations, only 9 were missense, with 107 
nonsense, 69 frameshift, and 15 splice-site mutations likely leading to 
haploinsufficiency at the cellular level. An additional 72 variations identified 
in the 642 tested samples (11%) were considered to have unknown clinical 
significance. Copy number changes (deletion/duplication of the entire gene or 
one/several exons) were found to account for a very small number of cases (n = 
3). This cohort represents the largest CHARGE syndrome sample size to date and 
is intended to serve as a resource for clinicians, genetic counselors, 
researchers, and other diagnostic laboratories.