May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Development of a Mutation Panel for Screening Mutations in Pediatric Patients With Abnormal Vasculogenisis
Author Affiliations & Notes
  • W. Dailey
    Beaumont Eye Institute, William Beaumont Hospital, Royal Oak, MI
  • K. Drenser
    Beaumont Eye Institute, William Beaumont Hospital, Royal Oak, MI
  • P. Doshi
    Beaumont Eye Institute, William Beaumont Hospital, Royal Oak, MI
  • M. Trese
    Beaumont Eye Institute, William Beaumont Hospital, Royal Oak, MI
  • Footnotes
    Commercial Relationships  W. Dailey, Beckman Coulter, F; K. Drenser, Beckman Coulter, F; P. Doshi, None; M. Trese, None.
  • Footnotes
    Support  Beckman Coulter Corporation
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 4603. doi:
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      W. Dailey, K. Drenser, P. Doshi, M. Trese; Development of a Mutation Panel for Screening Mutations in Pediatric Patients With Abnormal Vasculogenisis . Invest. Ophthalmol. Vis. Sci. 2006;47(13):4603.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Abnormal retinal vasculature is present in patients with various pediatric vitreoretinal diseases such as FEVR, Norrie’s, PFVS and ROP. We collected genomic DNA from 160 patients with pediatric retinal diseases(FEVR;n=32, Norries;n=3, ROP;n=96). Three genes (NDP, Fz4 and LRP–5) of the Wnt ß–cantenin signaling pathway have been implicated in vascularization of the retina. A panel of known mutations was selected for these three genes. Appropriate primers were designed and a protocol was optimized to efficiently and accurately screen the selected panel of mutations.

Methods: : Single base extension technology on a capillary electrophoreses platform was used to screen mutations using genomic DNA. Initially, single mutations were tested individually. PCR and interrogation primers were then designed to construct a multiplex panel. Primer/template concentrations, thermocycling and cleanup conditions were optimized to obtain well separated products. The reactions were separated and analyzed using the Beckman CEQ 8000 genetic analysis system.

Results: : The mutation panel was successfully developed. This assay yields reproducible and accurate results.

Conclusions: : This Beckman Coulter SNP kit provides and easy and efficient means for screening three genes of the Wnt ß–cantenin signaling pathway.

Keywords: mutations • retinal neovascularization • gene screening 
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