Detection of four different 11q23 chromosomal abnormalities by multiplex-PCR and fluorescence-based automatic DNA-fragment analysis.

摘要: A nested polymerase chain reaction (PCR) protocol was developed for rapid detection of four different 11q23 abnormalities by a single PCR assay. During each the two rounds sense primer located within exon 5 MLL gene at combined with antisense primers, possible translocation partner genes chromosomes 4, 6, 9, and 19, respectively. Except all primers used during second round nested-PCR carried characteristic fluorescence label their 5'-end. Agarose gel analysis products sufficient to discriminate between absence any rearrangements presence least one them. Discrimination not agarose due molecular heterogeneity breakpoints resulting in variable size. For this reason, automatic fluorescence-based DNA-fragment exactly define if positive result had been obtained analysis. In patients leukemia, assay may enable fast highly sensitive abnormalities, which usually correlate poor clinical prognosis.