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Screening for the two most frequent mutations in Leber's hereditary optic neuropathy by duplex PCR based on allele‐specific amplification

This report describes a rapid and inexpensive assay, which allows detection, in whole blood and by PCR alone, of the two most frequent mitochondrial DNA mutations causing Leber's hereditary optic neuropathy. The assay is based on allele‐specific amplification, using primers with the mutation‐specific base in the 3′ position, and a deliberately introduced G→C Substitution of base no. four from the 3′ end, which prevents amplification of the wild‐type allele. © 1993 Wiley‐Liss, Inc.