The advent of BRAF-targeted therapies led to increased survival in patients with metastatic melanomas harboring a BRAF V600 mutation (implicated in 46-48% of malignant melanomas). The Idylla(™) System (Idylla(™)), i.e., the real-time-PCR-based Idylla(™) BRAF Mutation Test performed on the fully-automated Idylla(™) platform, enables detection of the most frequent BRAF V600 mutations (V600E/E2/D, V600K/R/M) in tumor material within approximately 90 min and with 1% detection limit. Idylla(™) performance was determined in a multi-center study by analyzing BRAF mutational status of 148 archival formalin-fixed paraffin-embedded (FFPE) tumor samples from malignant melanoma patients, and comparing Idylla(™) results with assessments made by commercial or in-house routine diagnostic methods. Of the 148 samples analyzed, Idylla(™) initially recorded 7 insufficient DNA input calls and 15 results discordant with routine method results. Further analysis learned that the quality of 8 samples was insufficient for Idylla(™) testing, 1 sample had an invalid routine test result, and Idylla(™) results were confirmed in 10 samples. Hence, Idylla(™) identified all mutations present, including 7 not identified by routine methods. Idylla(™) enables fully automated BRAF V600 testing directly on FFPE tumor tissue with increased sensitivity, ease-of-use, and much shorter turnaround time compared to existing diagnostic tests, making it a tool for rapid, simple and highly reliable analysis of therapeutically relevant BRAF mutations, in particular for diagnostic units without molecular expertise and infrastructure.
One‐hundred and twenty‐four amplified fragment length polymorphism (AFLP) and 49 random amplified polymorphic DNA (RAPD) markers have been used to distinguish between 20 and 23 commercial chicory cultivars, respectively. These were all Cichorium intybus var. foliosum F1 hybrids, currently used in hydroponic forcing. Five‐hundred and twenty RAPD primers (OPERON) were tested, of which 156 resulted in reproducible patterns and 26 yielded polymorphisms. Two‐hundred and fifty‐six AFLP primer‐combinations were tested and six combinations were selected for identification purposes. Similarity indices were measured and clustering has been done using pairwise comparison. Both types of marker provide similar conclusions. Two major clusters are formed, representing late and early cultivars. All cultivars were identified using 10 informative RAPD primers or three AFLP primer combinations. A low degree of polymorphism was detected between some early cultivars, suggesting a narrow genetic base in their breeding strategy.
The phylogenetic relationship between different Cichorium intybus cultivars and cultivar groups was studied. A total of 127 random amplified polymorphic DNA markers have been used to distinguish between 51 Cichorium intybus cultivars belonging to three different cultivar groups. A principal component analysis was performed on the complete data set and a clear grouping of chicory cultivar groups was observed. The three chicory groups tested (root chicory, witloof and pain de sucre) were separated from one another in the principal component plots. The first two axes accounted for 43.9% of the total variation. Although the chicory used in traditional forcing in soil and the red chicory cultivars were related more to each other and to the chicory cultivars from the witloof group, they still formed separate groups in the principal component plot.
The genetic basis of pith characteristics in chicory (Cichorium intybus L. var. foliosum Hegi) was investigated. Quantitative trait loci (QTL) were mapped in an F2 population (565 F2 plants) derived from a cross between two inbred chicory lines. A molecular marker linkage map of this cross had previously been constructed based on 129 random amplified polymorphic DNA markers. Each F2 plant was selfed and plant characteristics were measured in the F3 populations. Although variation in pith characteristics was largely environmentally influenced, QTL for the characteristics length of pith, browning of the pith, hollow pith and apple pith were detected in many linkage groups. Interactions between QTL were found for the three characteristics: pith length, browning of the pith and hollow pith. The QTL detected confirmed the early forcing suitability of the one parent inbred line and late forcing suitability of the other.
Productivity is the main objective in chicory breeding, in addition to uniformity and reproducibility. The genetic bases of chicory (Cichorium intybus L. var. foliosum Hegi) production characteristics have been investigated in this paper. Characteristics were measured on F3 populations and quantitative trait loci (QTLs) were mapped on to the genetic linkage map of chicory. A large part of the total variation was explained by the QTLs detected in the first forcing period. In the winter and late forcing period, less variation was explained. QTLs were also found for the taste characteristics of bitterness and sweetness in the fresh non-cooked chicon.
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