In-house production of Fluorescence in situ hybridization probes for alpha-satellite centromeric region for detection of chromosomal aneuploidy

Abstract

Fluorescence in situ hybridization (FISH) depends on the complementary nature of the labeled probe and its targeted chromosomal segment. FISH technique allows the detection of numerical chromosomal aneuploidy by using the centromeric probes. Numerical aberrations represent a significant percentage of chromosomal abnormalities and is a main cause of pregnancy loses and cancer. The present study aimed to produce in-house FISH centromeric probes that help in the identification of numerical chromosomal aberrations (copy number variants) and can be used as a control probe. In this study, the probes were produced from human genomic DNA using Polymerase Chain Reaction (PCR) with specific primers. PCR was used for amplification and labeling of alpha satellite centromeric probes of chromosomes 1 and 7. The alpha satellite centromeric probes allow us to identify the numerical abnormalities of its corresponding chromosome. All the produced probes were sensitive and specific for the detection of chromosomal copy number variants. To our knowledge this is the first time to produce FISH probes in Egypt; our next plan is to produce double color probes for specific locus.