Scanning detection of mutations in human ornithine transcarbamoylase by chemical mismatch cleavage

M. Grompe, D. M. Muzny, C. T. Caskey

Research output: Contribution to journalArticle

102 Scopus citations

Abstract

The mutations in X chromosome-linked diseases are frequently heterogeneous, and yet only a small fraction can be detected by Southern analysis. We therefore adapted the chemical cleavage method of Cotton et al. [Cotton, R.G.H., Rodrigues, N.R. and Campbell, R.D. (1988) Proc. Natl. Acad. Sci. USA 85, 4397-4401] and the polymerase chain reaction to rapidly scan for point mutations in X chromosome-linked ornithine transcarbamoylase (carbamoyl-phosphate: L-ornithine carbamoyltransferase, EC 2.1.3.3) deficiency. This simple heteroduplex mapping method identified different mismatch sites in polymerase chain reaction-amplified liver cDNA from five unrelated ornithine transcarbamoylase-deficient patients. The predicted sequence alteration was confirmed by DNA sequencing in all five patients and indicated a likely disease-causing mutation in four of these patients. In one atypical ornithine transcarbamoylase-deficient patient a sequence alteration compatible with a cDNA polymorphism was found. One family was studied in detail. Female-carrier detection was performed by chemical cleavage of amplified genomic DNA and verified by allele-specific oligonucleotide hybridization. This mutation scanning approach is simple, sensitive, and applicable to many genetic loci.

Original languageEnglish (US)
Pages (from-to)5888-5892
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume86
Issue number15
DOIs
StatePublished - 1989

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