Preimplantation Genetic Diagnosis of X-linked Severe Combined Immunodeficiency
- VernacularTitle:X-连锁严重联合免疫缺陷病的植入前遗传学诊断
- Author:
tao Hai WU
1
;
ting Xiao SHEN
;
jian Qing YE
;
liang Yu LIU
;
ping Yi ZHONG
;
hong Yan ZENG
;
wen Yan XU
;
quan Can ZHOU
Author Information
1. 中山大学附属第一医院生殖医学中心
- Keywords:
X-linked severe combined immunodeficiency;
preimplantation genetic diagnosis(PGD);
multiple displacement amplifi?cation(MDA);
short tandem repeat (STR);
haplotype analysis
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2017;38(6):955-960
- CountryChina
- Language:Chinese
-
Abstract:
[Objective]To establish a reliable and accurate preimplantation genetic diagnosis (PGD)method using multiple dis?placement amplification (MDA), which can be applied to the diagnosis of X-linked severe combined immunodeficiency disease (X-SCID).[Methods]Haplotype analysis for the X-SCID family was performedusing five short tandem repeats (STR) markers flanking the both sides of the interleukin-2 (IL-2) receptor gamma chain (IL2RG) gene. MDA technique was used for single-cell whole genomic amplification. The products were used as template in polymerase chain reaction (PCR) of informative STR markers found by linkage analysis for haplotype analysis as well as sequencing of the IL2RG gene exon 5.The amelogenin (AMEL) locus was used to do sex diag?nosis.[Results]Linked analysis revealed 3 STR markers were informative. The method was evaluated with 10 single lymphocytes and 10 single blastomeres. MDA was successful in all single cell. The detection efficiency of gene sequencing of pathogenic IL 2RG exon5 was 100%. The PCR efficiency of 3 STR informative markers and AMEL was 96.3%(77/80)and the average allele drop-out (ADO) rate was 11.5%(7/61). A cycle of PGD was performed on the family, and seven embryos were diagnosed, two of which were normal embry?os. Twin pregnancy occurred after transplantation which were given a healthy baby boy and a healthy baby girl.[Conclusion]In this study, multiple displacement amplification combined with specific amplification/sequencing of pathogenic gene and haplotype analysis in the single cell level of X-linked severe combined immunodeficiency disease were performed. The protocol can avoid misdiagnosis caused by contamination and ADO, and improve the diagnostic efficiency of PGD.
