SMA carrier testing - validation of hemizygous SMN exon 7 deletion test for the identification of proximal spinal muscular atrophy carriers and patients with a single allele deletion
H. Scheffer et al., SMA carrier testing - validation of hemizygous SMN exon 7 deletion test for the identification of proximal spinal muscular atrophy carriers and patients with a single allele deletion, EUR J HUM G, 8(2), 2000, pp. 79-86
To facilitate the detection of carriers of a hemizygous survival motor neur
on (SMN) exon 7 deletion we have modified the quantitative SMN exon 7 assay
described by McAndrew ct al (1997). The major changes include quantitative
analysis of the amount of SMN exon 7-specific fluorescently-labelled PCR p
roduct on an automated sequencer, and the monitoring of the completeness of
a Dral digestion necessary to distinguish the PCR products of exons 7 of S
MN and its copy gene. In our method the amount of SMN exon 7 PCR product is
compared with the amount of a co-amplified PCR product of the retinoblasto
ma (RB7) exon containing a Dral restriction site. By co-amplification using
the same primers of plasmids included in the reaction as internal standard
s containing SMN exon 7 with a 36-nucleotide deletion and RB7 exon 13 with
a 19-nucleotide deletion, respectively, the relative amplification efficacy
can be monitored. The assay has been validated in 63 ascertained carriers
and 28 ascertained non-carriers. The sensitivity of the test is approximate
ly 97%, the specificity approaches 100%. In four out of six SMA patients wi
thout a homozygous deletion we detected a hemizygous deletion. The implicat
ions of the use of this assay for carrier testing and for confirmation of t
he clinical diagnosis of SMA are discussed.