Presented at the 32nd Association for Research in Otolaryngology MidWinter Meeting, Baltimore, Maryland, U.S.A., February 14–19, 2009.
Application of SNaPshot multiplex assays for simultaneous multigene mutation screening in patients with idiopathic sensorineural hearing impairment†
Article first published online: 28 AUG 2009
Copyright © 2009 The American Laryngological, Rhinological, and Otological Society, Inc.
Volume 119, Issue 12, pages 2411–2416, December 2009
How to Cite
Wu, C.-C., Lu, Y.-C., Chen, P.-J., Liu, A. Y.-Z., Hwu, W.-L. and Hsu, C.-J. (2009), Application of SNaPshot multiplex assays for simultaneous multigene mutation screening in patients with idiopathic sensorineural hearing impairment. The Laryngoscope, 119: 2411–2416. doi: 10.1002/lary.20621
- Issue published online: 20 NOV 2009
- Article first published online: 28 AUG 2009
- Manuscript Accepted: 9 JUN 2009
- National Science Council of the Executive Yuan of the Republic of China. Grant Number: 97-2314-B-002-092-MY3
- SNaPshot technique;
- mitochondrial 12S rRNA;
- genetic test
To develop a cost-effective and robust genetic diagnostic tool for patients with idiopathic nonsyndromic sensorineural hearing impairment.
Development of a diagnostic tool and validation in a prospective cohort.
Twenty common sequence variants in GJB2, SLC26A4, and the mitochondrial 12S rRNA gene were selected based on our previous epidemiological study. These variants were analyzed using the SNaPshot technique. The efficacies of the SNaPshot multiplex assays were determined by using a prospective cohort composed of 214 unrelated Taiwanese patients with idiopathic sensorineural hearing impairment. The results of the assays were compared to the results obtained by direct sequencing.
We developed a diagnostic technique consisting of two consecutive panels of SNaPshot multiplex assays, with each panel screening 10 common sequence variants. Theoretically, this design can detect more than 98% of the known deafness-associated sequence variants in Taiwanese individuals. A total of 126 (58.9%) patients were diagnosed as having at least one sequence variant using the SNaPshot multiplex assays. In total, the SNaPshot assays yielded an accuracy of more than 99%.
The strengths of SNaPshot multiplex assays include high accuracy, high sensitivity, high flexibility (the examination panel can be easily expanded for additional mutations), low cost (less than US $10 per patient), and easy implementation for any institute with a DNA sequencer. Although only 20 to 30 mutations can be examined in two to three runs of the SNaPshot assay, this technology may be suitable for first-pass screening of deafness-associated mutations in populations with a relatively homogeneous ethnic background. Laryngoscope, 2009