Comparative study of Multiplex Ligation - dependent Probe Amplification (MLPA) and Microsatellite assays to detect chromosomes 1p36 and 11q loss of heterrozygosity in Neuroblastoma

Abstract

Neuroblastoma (NB) is the common solid tumor occurring in childhood. There are several genetic factors affect the NB classification such as MYCN amplification, deletion on chromosome 1 and 11, gain of chromosome 17. NB were divided into three main groups which were High- risk, Intermediate and Low- risk. Classification group helps find out the suitable treatment for patients that lead to increase survive ability. Patient with MYCN amplification is classified into High- risk group with poor prognostic. Loss of heterozygosity on short arm of chromosome 1 (1p36 LOH) and long arm of chromosome 11 (11q LOH) are important genetic aberration that impact distribution group in NB. Patient with 1p LOH or 11q LOH is classified into Intermediate group. There are several techniques that can be used for chromosome deletion such as G- banding, fluorescence in situ hybridization technique (FISH), Comparative genomic hybridization (CGH). However, all of them have some limitations. Microsatellite and MLPA are effective assays that easily establish in Vietnam laboratory. In this study, we found out some Microsatellite markers which have matched position with typical genes that have high frequent occur in NB base on MLPA kit. 19 tumors and their matched blood samples were investigated randomly for detection LOH on chromosome 1(5 primers) and 11(8 primers) by both Microsatellite and MLPA assays. Comparative analysis is tested by kappa index values showing moderate concordance between two techniques in detecting 1p LOH and 11q LOH. Results from 2 methods also were validated with previous research and result of MYCN amplification to increase accuracy. The sensitivity and specificity are interested. We found out that MLPA is high accuracy, sensitivity and specificity than Microsatellite. In summary, MLPA is a rapid, reliable and high- throughput method for the detection of chromosomes 1p36 and 11q loss of heterozygosity in Neuroblastoma. Keyword: Neuroblastoma, loss of heterozygosity, 1p36 LOH, 11q LOH, Microsatellite, MLPA