%0 Journal Article %J Diagn Mol Pathol %D 2011 %T High-throughput genotyping robot-assisted method for mutation detection in patients with hypertrophic cardiomyopathy. %A Bortot, Barbara %A Athanasakis, Emmanouil %A Brun, Francesca %A Rizzotti, Diego %A Mestroni, Luisa %A Sinagra, Gianfranco %A Severini, Giovanni Maria %K Cardiomyopathy, Hypertrophic %K DNA Mutational Analysis %K Genetic Predisposition to Disease %K Genetic Testing %K Genotyping Techniques %K High-Throughput Nucleotide Sequencing %K Humans %K Muscle Proteins %K Mutation %K Robotics %X

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.

%B Diagn Mol Pathol %V 20 %P 175-9 %8 2011 Sep %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/21817903?dopt=Abstract %R 10.1097/PDM.0b013e31820b34fb