High-throughput genotyping robot-assisted method for mutation detection in patients with hypertrophic cardiomyopathy

Barbara Bortot, Emmanouil Athanasakis, Francesca Brun, Diego Rizzotti, Luisa Mestroni, Gianfranco Sinagra, Giovanni Maria Severini

Research output: Contribution to journalArticlepeer-review


Hypertrophic cardiomyopathy (HCM) is the most frequent autosomal dominant genetic heart muscle disease and the most common cause of sudden cardiac death in young people (under 30 y of age), who are often unaware of their underlying condition. Genetic screening is now considered a fundamental tool for clinical management of HCM families. However, the high genetic heterogeneity of HCM makes genetic screening very expensive. Here, we propose a new high-throughput genotyping method based on a HCM 96-well sequencing plate for the analysis of 8 of the most frequent HCM-causing sarcomeric genes by automating several processes required for direct sequencing, using a commercially available robotic systems and routinely used instruments. To assess the efficiency of the robot-assisted method, we have analyzed the entire coding sequence and flanking intronic sequences of the 8 sarcomeric genes in samples from 18 patients affected by HCM and their relatives, which revealed 9 different mutations, 3 of which were novel. The automated, robot-assisted assembling of polymerase chain reaction, purification of polymerase chain reaction products, and assembly of sequencing reactions resulted in a substantial saving of time, reagent costs, and reduction of human errors, and can therefore be proposed as a primary strategy for mutation identification in HCM genetic screening in many medical genetic laboratories.

Original languageEnglish
Pages (from-to)175-179
Number of pages5
JournalDiagnostic Molecular Pathology
Issue number3
Publication statusPublished - Sep 2011


  • genetic screening
  • hypertrophic cardiomyopathy
  • mutation
  • robotic
  • sarcomeric gene

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Cell Biology
  • Molecular Biology


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