Development and implementation of a novel panel consisting 16 str markers linked to the hla-gene-cluster applicable for hla-typing, pnd&pgd

Kiyana alsadat Fatemi,1 Maryam abiri,2 Zohreh sharifi,3 Solmaz sabeghi,4 Hanieh noferesti,5,*

1. 1. Dr. Zenati’s Medical Genetics Laboratory, Kawsar Human Genetics Research Center, Tehran, Iran
2. 3. Department of Medical Genetics and Molecular Biology, School of Medicine, Iran University of Medical Sciences, Tehran
3. 1. Dr. Zenati’s Medical Genetics Laboratory, Kawsar Human Genetics Research Center, Tehran, Iran
4. 1. Dr. Zenati’s Medical Genetics Laboratory, Kawsar Human Genetics Research Center, Tehran, Iran
5. 1. Dr. Zenati’s Medical Genetics Laboratory, Kawsar Human Genetics Research Center, Tehran, Iran

Abstract

Introduction

Pgd (pre-implantation genetic diagnosis) in combination with human leukocyte antigen (hla) matching, allow not only selection and transfer of unaffected embryos but also have a close hla match with the existing affected child. in these cases, pgd not only used to avoid the birth of affected children but also to conceive healthy children, who can be a potential hla-identical donors of haematopoietic stem cells (hsc) for transplantation in affected siblings. this study aimed to develop a novel panel containing 16 polymorphic str (short tandem repeat) markers linked to the hla gene clusters for use in hla typing and in pgd for different diseases. it can be also used for the combination of pgd and hla typing together.

Methods

Polymorphic str markers were selected from tandem repeats finder and sequence-based estimation of repeat variability databases. suitable primers were designed to setup a multiplex-pcr for hla haplotyping . 50 unrelated individuals were genotyped to assess the allele frequencies, heterozygosity of the selected markers. genotyping of each individual were performed using fragment analysis. statistical analysis was performed using genalex6.03.

Results

Our results showed that the heterozygosity of selected markers were between38%-89%. totally 140 different alleles were observed for 16 loci, with the frequency of 0.012-0.413. the most number of alleles were seen in 3s320 and the lowest was found in cs366.3 loci. also the other loci had acceptable polymorphism information content.

Conclusion

All studied markers were in hardy-weinberg equilibrium. the panel provides a powerful and reliable molecular hla typing method applicable in pgd.

Keywords

Str,hla,haplotype, panel,pgd,pnd