Using two genetic tests in sequence can substantially improve the detection of Duchenne muscular dystrophy (DMD), especially when initial results are negative, according to a study published recently in PLOS One.
Evidence suggests that starting with multiplex ligation-dependent amplification (MLPA) — a method that detects DNA insertions and deletions — and following with next-generation sequencing (NGS) to identify small mutations in the DMD gene increases the likelihood of confirming a diagnosis, which may help patients and families access appropriate care sooner.
“These results are consistent with the recommendations of international guidelines on the molecular genetic approach in patients with DMD/[Becker muscular dystrophy],” explained the study’s authors.
A meta-analysis of studies published through July 2025 evaluated how well MLPA and NGS identified DMD or Becker muscular dystrophy (BMD) in patients who doctors suspected might have one of the conditions. Researchers reviewed 10 MLPA studies including 3,786 patients and 14 NGS studies including 4,333 patients. These studies compared genetic testing results with established reference standards such as biopsy or sequencing.
Read more about DMD testing and diagnosis
MLPA demonstrated strong performance: MLPA correctly identified most patients with DMD and has a relatively low rate of false positives. However, it does not detect all cases, particularly those caused by smaller genetic changes.
NGS, which can identify more subtle mutations, was able to correctly detect 77% of cases. Importantly, when NGS was used after a negative MLPA result, the detection rate increased to 97%. In a hypothetical group of 1,000 patients, this combined approach could correctly diagnose 931 individuals while misclassifying only 29. By comparison, MLPA or NGS alone resulted in more missed or incorrect diagnoses.
For patients, this stepwise testing strategy may reduce uncertainty and shorten the time to diagnosis. Earlier and more accurate identification of DMD can guide treatment decisions, genetic counseling and eligibility for clinical trials. However, there are trade-offs. MLPA is faster (10 to 12 days) and less expensive, while NGS can take two to eight weeks and cost several times more. Access and cost may influence which tests are used.
Although the findings support combined testing, the certainty of evidence remains low due to variability across studies. Still, for patients with strong clinical signs but negative initial results, adding a second test may provide critical answers and improve care planning.
