title: Development and Validation of a Mass Spectrometry-Based Assay for the Molecular Diagnosis of Mucin-1 Kidney Disease.
abstract: Mucin-1 kidney disease, previously described as medullary cystic kidney disease type 1 (MCKD1, OMIM 174000), is an autosomal dominant tubulointerstitial kidney disease recently shown to be caused by a single-base insertion within the variable number tandem repeat region of the MUC1 gene. Because of variable age of disease onset and often subtle signs and symptoms, clinical diagnosis of mucin-1 kidney disease and differentiation from other forms of hereditary kidney disease have been difficult. The causal insertion resides in a variable number tandem repeat region with high GC content, which has made detection by standard next-generation sequencing impossible to date. The inherently difficult nature of this mutation required an alternative method for routine detection and clinical diagnosis of the disease. We therefore developed and validated a mass spectrometry-based probe extension assay with a series of internal controls to detect the insertion event using 24 previously characterized positive samples from patients with mucin-1 kidney disease and 24 control samples known to be wild type for the variant. Validation results indicate an accurate and reliable test for clinically establishing the molecular diagnosis of mucin-1 kidney disease with 100% sensitivity and specificity across 275 tests called.
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts. Electronic address: brendan@broadinstitute.org.
authors.forename: Brendan
authors.initials: B
authors.lastname: Blumenstiel
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts.
authors.forename: Matthew
authors.initials: M
authors.lastname: DeFelice
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts; Partners Healthcare Laboratory for Molecular Medicine, Cambridge, Massachusetts.
authors.forename: Ozge
authors.initials: O
authors.lastname: Birsoy
authors.affiliation: Nephrology Section, Wake Forest Baptist Medical Center, Winston-Salem, North Carolina.
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Bleyer
authors.affiliation: Institute for Inherited Metabolic Disorders, Charles University in Prague, Prague, Czech Republic.
authors.forename: Stanislav
authors.initials: S
authors.lastname: Kmoch
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts.
authors.forename: Todd A
authors.initials: TA
authors.lastname: Carter
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts.
authors.forename: Andreas
authors.initials: A
authors.lastname: Gnirke
authors.affiliation: Nephrology Section, Wake Forest Baptist Medical Center, Winston-Salem, North Carolina.
authors.forename: Kendrah
authors.initials: K
authors.lastname: Kidd
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts; Partners Healthcare Laboratory for Molecular Medicine, Cambridge, Massachusetts.
authors.forename: Heidi L
authors.initials: HL
authors.lastname: Rehm
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts.
authors.forename: Lucienne
authors.initials: L
authors.lastname: Ronco
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts.
authors.forename: Eric S
authors.initials: ES
authors.lastname: Lander
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts.
authors.forename: Stacey
authors.initials: S
authors.lastname: Gabriel
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts.
authors.forename: Niall J
authors.initials: NJ
authors.lastname: Lennon
mesh_terms: D005838:Genotype; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D025202:Molecular Diagnostic Techniques; D018396:Mucin-1 / Q000235:genetics*; D009154:Mutation; D016891:Polycystic Kidney, Autosomal Dominant / Q000175:diagnosis* / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D057188:Workflow
chemical_list: D018396:Mucin-1