Microdissection based comparative genomic hybridization analysis (micro-CGH) of secondary acute myelogenous leukemias

Comparative genomic hybridization (CGH) is a well established technique in molecular cytogenetics. However, leukemias, and especially secondary acute myelogenous leukemias (sAML) are not very well analyzed by this technique, even though such diseases are often characterized by complex karyotypic cha nges, not resolvable by conventional cytogenetic banding analysis. This lac k of CGH-studies might be due to the fact, that in most cases bone marrow a spirate is too limited to do DNA-extraction additionally to the cytogenetic analysis. To circumvent this problem a new CGH technique has been applied to analyze 10 AML cases with complex karyotypic changes. In each case 15 in terphase nuclei of the harvested and fixed bone marrow cell-suspension have been microdissected from the coverslip surface and collected in a tube. Su bsequently, DNA was amplified by DOP-PCR. With this micro-CGH technique add itional cytogenetic information from 10 highly aberrant AML cases was obtai ned and confirmed by FISH on metaphase of the corresponding AML case.