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Immunohistochemistry and Reverse Transcriptase–Polymerase Chain Reaction as Methods for Diagnostic Determination of Usher Syndrome Type IIa
Author(s) -
Cohn Edward,
Bhattacharya Gautam,
Pearsall Nicole,
Shendrik Igor,
Kimberling William,
Cosgrove Dominic
Publication year - 2004
Publication title -
the laryngoscope
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.181
H-Index - 148
eISSN - 1531-4995
pISSN - 0023-852X
DOI - 10.1097/00005537-200407000-00032
Subject(s) - immunostaining , immunohistochemistry , salivary gland , pathology , stain , biology , biopsy , type iv collagen , basement membrane , staining , antibody , saliva , microbiology and biotechnology , medicine , immunology , extracellular matrix , laminin , biochemistry
Objectives/Hypothesis: 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. Study Design: 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. Conclusion: The feasibility was confirmed of diagnosing Usher syndrome type IIa using purified usherin antibody in subjects having two null USH2A mutations.