Immunohistochemistry and reverse transcriptase-polymerase chain reaction as methods for diagnostic determination of usher syndrome type IIa.
OBJECTIVE
Patients having null mutations in the USH2A gene do not produce usherin and therefore are not positive for immunohistochemical staining of the usherin protein. Thus, immunostaining for usherin can serve as a reliable diagnostic tool for Usher syndrome type IIa.


METHODS
Prospective.


METHODS
Immunohistochemical staining for usherin was carried out in basement membrane of minor salivary gland tissue from subjects with confirmed Usher syndrome type IIa and from archival minor salivary gland tissue from patients without Usher syndrome as control samples. Quantitative usherin messenger RNA analysis was performed using minor salivary gland biopsy tissue.


RESULTS
Five subjects with Usher syndrome type IIa had no immunostaining in minor salivary gland tissue, whereas control minor salivary gland tissue did stain with usherin antibody. No usherin RNA was detected in biopsy specimens from patients with confirmed Usher syndrome IIa.


CONCLUSIONS
The feasibility was confirmed of diagnosing Usher syndrome type IIa using purified usherin antibody in subjects having two null USH2A mutations.
Cohn
Edward
E
Usher Syndrome Center, Boys Town National Research Hospital, Omaha, NE 68131, USA.
Bhattacharya
Gautam
G
Pearsall
Nicole
N
Shendrik
Igor
I
Kimberling
William
W
Cosgrove
Dominic
D
D001485:Basement Membrane; D003638:Deafness / Q000175:diagnosis*; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008854:Microscopy, Electron; D009154:Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012469:Salivary Glands / Q000378:metabolism / Q000473:pathology; D013577:Syndrome
D016326:Extracellular Matrix Proteins; C112735:USH2A protein, human