A study of micro-satellite assay to detect the chromosome 1P36 loss in neuroblastoma

Abstract

Neuroblastoma (NB) is pediatric cancer that arises from precursor cells of mature dorsal ganglion neuron which often lead to abnormal proliferative state in young children and infants (younger than 2 years of age). NB is heterogeneous phenotypes therefore it is hard to manage effectively in diagnostic clinics. It is mainly dependent on laborious and time-consuming histology's evaluation. Aside from MYC-N copy status instability of chromosome 1poftenlead to aggressive disease in NB patients. Both of them can be identified by G-band chromosome technique. In this study, I developed a molecular based PCR-technique to identify the loss of somatic 1p36 by using six specific micro-satellite markers for NB. The 8 tumors and its matched-germlines (blood) is assigned to run on a precise 8-10% polyacrylamide gel based on the appearance of specific, size amplicons (PCR products) corresponding to the target micro-satellites. The results showed the optimization of different genomic DNA isolation, PCR set-up, and polyacrylamide gel electrophoresis for the precise identification of small base pair fractions of micro-satellites. Moreover, the elimination of PCR carry-over contamination and erroneous reporting of false-positives were confirmed by CGH-array. Overall, the micro-satellites described in this study are specific for the routine detection of loss of 1p36 in NB samples. Key words: neuroblastoma, 1p36 chromosome loss, micro-satellite markers, PCR.