Table 2.
Techniques currently used in diagnosis of hypertrophic cardiomyopathy: major providers, advantages, and disadvantages
| Technique | Major providers | Advantages | Disadvantages | References |
|---|---|---|---|---|
| Sanger sequencing | Thermo Fisher Scientific | Accurate Precise Important for validation/confirmation proposes |
Time-consuming (more than 1 month; totaltime depends on number of regions analyzed) Laborious Expensive |
* |
| dHPLC | Transgenomic, Inc. | Faster than Sanger sequencing (total time depends on number ofregions analyzed) | Sensitivity and specificity 87.5% and 97.42%,respectively (compared with Sanger sequencing) Optimization laborious Screening of heterozygous/homozygous Require Sanger sequencing for mutation confirmation |
61,67,104–106 |
| High resolution melting | Hoffman-La Roche Ltd. Qiagen NV Thermo Fisher Scientific Qiagen NV |
>99% sensitivity and specificity (compared with Sanger sequencing) Closed-tube genotyping approach, reducing contamination risk Much faster than Sanger sequencing; 3 hours for complete genescanning (96 or 384 plates; total time depends on number of regions analyzed) |
Require Sanger sequencing for mutation confirmation | 46,56,61,107 |
| iPLEX-MassARRAY | Sequenom, Inc. | 48 hours for complete mutation detection (multiplex analysis of morethan 10,000 genotypes/384 plate) Accurate |
Incapable to detect unknown mutations | 56,75,108 |
| Next-generation sequencing | Illumina, Inc. Thermo Fisher Scientific Hoffman-La Roche Ltd. |
Allow simultaneous sequencing of large amount of genes Targeted gene panels – limited gene panels allows high depth ofcoverage for increased sensitivity and specificity Higher ability to interpret the findings in a clinical context Allows to run more patient samples per instrument cycle (barcoding and pooling) Less amount of data and storage requirements |
Genome data analysis is time-consuming High number of VUS Follow-up by Sanger sequencing to fill gaps in the datafor regions showing low coverage (eg, GC-rich orrepetitive regions) |
53,60,62,76, 81–88 |
Note:
*
Sanger sequencing is used in all the references provided in this table as the confirmation/validation gold standard technique.
Abbreviations: dHPLC, denaturing high-performance liquid chromatography; VUS, variants of uncertain significance.