For pathogenicity assessment, mycelia cultured in CM liquid medium for 2 days were harvested (5,000×g, 2 min), washed twice with distilled water, and fragmented into the length of 30 50 µm using a homogenizer.
The pathogenicity assessment was performed using the injection method on sixth-instar G. mellonella larvae.
On the other hand, when performing extensive mutation analyses, problems with pathogenicity assessment can occur for some types of mutations.
However, in silico predictions are not always consistent with functional studies and, despite recent advances, pathogenicity assessment remains challenging, particularly for hypomorphic, synonymous and non-coding variants.
This finding is particularly important for genetic counselling in HNPCC, where the pathogenicity assessment of any nucleotide substitution is crucial to correctly predict cancer risk.
The verification of the model is indicated in terms of the amount of steps required for pathogenicity assessment of each VUS. Figure 2 demonstrates the collected results of each required assessment step for each gene and its variations.
Pathogenicity assessment is based on family co-segregation, functional predictions, the degree of conservation of amino-acids and also on the analysis of precise phenotypical descriptions [ 1, 13, 14].
In the past few years, the generation of fast, powerful, and cost-effective gene discovery and mutation detection methods, in addition to the development of variant pathogenicity assessment tools, has markedly increased the clinical utility of molecular diagnoses.
For nonsynonymous mutations, we assessed evolutionary conservation, and employed previously described pathogenicity score assessment tools.
This strategy ignores computational assessments of pathogenicity, although this situation could change if these tools became more predictive.
