Population-Wide Duchenne Muscular Dystrophy Carrier Detection by CK and Molecular Testing

Figure 4

Pedigrees (left) and the results of DMD gene testing (right). (a) Pedigree of the family of carrier 1 (III2). The carrier status is indicated by a circle with a dot, sloid symbols (affected), clear symbols (unaffected). The result of MLPA analysis on the right shows exon 44del in DMD gene (the horizontal axis represents exon number of DMD, and the vertical axis shows the relative peak area ratios compared to the mean of control samples). (b) Pedigree of the family of carrier 2 (I2). The carrier status is indicated by a circle with a dot, sloid symbols (affected), clear symbols (unaffected). The result of MLPA on the right shows exon 43del in DMD gene. (c) Pedigree of the family of carrier 3 (I2). The carrier status is indicated by a circle with a dot, sloid symbols (affected), clear symbols (unaffected). The result of MLPA on the right shows exon 52-54del in DMD gene. (d) A: pedigree of the family of carrier 4 (I2). The carrier status is indicated by a circle with a dot, abortion (a dot), clear symbols (unaffected). PCR-based Sanger sequencing on the right validates the results of next generation sequencing: the arrow represents the carrier (I2) with 1 heterozygous insertion variant c.10364 dup T of exon 73 (upper right) as well as the same but hemizygous variant in the aborted embryo (lower right). B: the amino acid sequences of the wild type and mutant were subjected to DNAMAN, revealing that the variant produced a truncated dystrophin. (e) Pedigree of the family of carrier 5 (I2). The carrier status is indicated by a circle with a dot, clear symbols (unaffected). The result of MLPA on the right shows exon 52_60dup in DMD gene. (f) A: pedigree of the family of carrier 6 (I2). The carrier status is indicated by circle with a dot, clear symbols (unaffected). PCR-based Sanger sequencing on the right validates the results of next generation sequencing: the arrow represents the carrier (I2) with 1 heterozygous missense variant c.7555G>A of exon 52. (b) Multiple species alignment analysis showed the high evolution conservation of amino acid sequence at the missense site.

(a)

(b)

(c)

(d)

(e)

(f)