Repeat-primed polymerase chain reaction in myotonic dystrophy type 2 testing

Myotonic dystrophy type 1 (DM1) and type 2 (DM2) are the most common autosomal dominant neuromuscular disorders in adults. DM1 is caused by an unstable expansion of the (CTG)(n) repeat tract in the DMPK gene, whereas DM2 is caused by an unstable expansion of the (CCTG)(n) repeat tract in the ZNF9 gene. The (CCTG)(n) repeat is a part of a complex repetitive motif (TG)(n)(TCTG)(n)(CCTG)(n), in which each of the elements is highly polymorphic. Repeat-primed polymerase chain reaction (PCR) is a commonly used technique for the determination of the presence or absence of the expanded alleles in both DM1 and DM2. Besides the expansion detection, it can be used for the determination of the repeat structure (repeat number, presence of interruptions, and their localization) in healthy-range alleles. Because the (CCTG)(n) part of the motif in DM2 is generally interrupted with other sequences, "tetraplet" repeat-primed PCR (TP-PCR) results interpretation is more complicated than for DM1. Most of the studies, published so far, used TP-PCR in a direction such that they amplified through the (TG)(n)(TCTG)(n) part of the motif. We compared the features of TP-PCR performed in the commonly used direction with the results obtained by TP-PCR performed in the opposite direction. Our results suggest that the direction that does not include the (TG)(n)(TCTG)(n) tract leads to better quality and more informative results in comparison with the direction containing the (TG)(n)(TCTG)(n) tract.