5. Automated analysis of 1p/19q status by FISH in oligodendroglial tumours.

Automated analysis of 1p and 19q status in oligodendroglial tumors by fluorescence in-situ hybridization (FISH) can be achieved by image-analysis software present in the majority of institutions using the FISH technique. Despite the widespread availability of this software, there are no specific guidelines in the literature on how to use it. We studied which green/red (G/R) probe signal combinations are predictive of 1p/19q co-deletion in a retrospective series of 53 oligodendroglial tumours and defined a new algorithm with a reduced sequence of combinations compared to previous studies. This algorithm was then tested and refined on a prospective series of 45 oligodendroglial tumours. The new algorithm scores 24 G/R combinations, which represent less than 50 % of the total observed combinations in our series. This algorithm excludes some previously described combinations and redefines the place of others. G/R combinations of 5/2, 6/2 and 6/3 associate with deletion status combinations, combinations of 1/2 associate with normal chromosome status, and combinations of 3/3 and 4/4 associate with imbalanced chromosome status. The new algorithm when applied to the combination and ratio methods of signal probe analysis gives a high concordance between manual and automated analysis on examination of 100 tumour cells (91% concordance for 1p and 89% concordance for 19q) and total concordance on examination of 200 tumour cells. This highlights the value of automated analysis to identify cases with imbalanced chromosome status, in which a larger number of tumour cells should be study by manual analysis. Our algorithm can be easily programmed on all existing FISH analysis software platforms and should facilitate multicentric evaluation and standardization of 1p/19q assessment in gliomas.