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

Hussein, Shymaa H; Amr, Khalda S; Abd El-Fattah, Abeer I; Mohamed, Amal M

doi:10.21608/aijpms.2022.121845.1114

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

Document Type : Original research articles

Authors

¹ Department of Human Cytogenetics, Institute of Human Genetics and Genome Research, National Research Centre, Cairo, Egypt

² Deparment of Medical Molecular Genetics, Institute of Human Genetics and Genome Research, National Research Centre, Cairo, Egypt

³ Department of Biochemistry, Faculty of Pharmacy (Girls), Al-Azhar University, Cairo, Egypt

10.21608/aijpms.2022.121845.1114

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 abnormalties 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 using Polymerase Chain Reaction (PCR) with specific primers and human genomic DNA. 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 colour probes for specific locus.

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