Diagnosis of Laron syndrome using monoplex-polymerase chain reaction technology with a whole-genome amplification template: A case report  

作　　者：Adina Neumann Miguel Angel Alcantara-Ortigoza Ariadna Gonzalez-del Angel Felipe Camargo-Diaz Esther Lopez-Bayghen 

机构地区：[1]Laboratorio de Investigacion y Diagnostico Molecular,Instituto de Infertilidad y Genetica Mexico SC,INGENES,Mexico City 05320,Mexico [2]Instituto Nacional de Pediatria,Torre de Investigacion,Mexico City 04530,Mexico [3]Laboratorio de Biologia Molecular,Departamento de Genetica Humana,Instituto Nacional de Pediatria,Mexico City 04530,Mexico [4]Departamento de Toxicologia,Centro de Investigacion y de Estudios Avanzados del Instituto Politecnico Nacional(CINVESTAV-IPN),Mexico City 07360,Mexico

出　　处：《World Journal of Clinical Cases》2019年第23期4029-4035,共7页世界临床病例杂志

基　　金：Conacyt,NO.231793

摘　　要：BACKGROUND Laron syndrome(LS)is an autosomal recessive hereditary condition affecting only 1/1000000 births.The cause is associated with mutations in the growth hormone(GH)receptor(GHR),leading to GH insensitivity.LS patients typically present with severe growth retardation,obesity,and abnormal sexual maturation.Currently,LS diagnosis is performed post-delivery.Therefore,we assessed the efficiency of Pre-implantation Genetic Testing(PGT)coupled with monoplex-polymerase chain reaction(PCR)technology for detecting this monogenic disease in embryos from a couple confirmed as LS heterozygous carriers CASE SUMMARY The couple LS-carriers were confirmed by the presence of a first child born with LS.The couple underwent a standard in vitro fertilization(IVF)protocol.DNA was collected from trophectoderm cells from day 5 embryos.Whole genome amplification(WGA)was performed using a Sureplex DNA Amplification System and analyzed by PCR,targeting the deletion of the exons 5 and 6 in the GHR gene as well as PGT by Next-generation Sequencing(Illumina).Eleven embryos were collected and analyzed.27.3%were the wild type for GHR,45.5%were heterozygotes,and 18.2%homozygous mutants.One embryo yielded no results.Three 2-embryos transfers were performed;2 normal homozygous and four heterozygous carriers were selected for transfer.The first two transfers were unsuccessful,whereas the final transfer with two heterozygous embryos resulted in clinical pregnancy.The genomic composition of the fetus was verified,applying the same techniques using amniocytes,extracted after 21 wk of the ongoing pregnancy.The fetus was confirmed as GHR deletion in exon 5-6,carrier.A non-affected baby was born.CONCLUSION Here,we present a case demonstrating that using WGA as a template in addition to PCR targeting specific gene regions,exons 5 and 6 on the GHR gene,could identify LS carrier embryos.This provides evidence that WGA and PCR serve as an excellent tool to detect this specific monogenic disease in IVF embryos,thus allowing selection of candidate embrBACKGROUND Laron syndrome(LS) is an autosomal recessive hereditary condition affecting only 1/1000000 births. The cause is associated with mutations in the growth hormone(GH) receptor(GHR), leading to GH insensitivity. LS patients typically present with severe growth retardation, obesity, and abnormal sexual maturation. Currently, LS diagnosis is performed post-delivery. Therefore, we assessed the efficiency of Pre-implantation Genetic Testing(PGT) coupled with monoplex-polymerase chain reaction(PCR) technology for detecting this monogenic disease in embryos from a couple confirmed as LS heterozygous carriers CASE SUMMARY The couple LS-carriers were confirmed by the presence of a first child born with LS. The couple underwent a standard in vitro fertilization(IVF) protocol. DNA was collected from trophectoderm cells from day 5 embryos. Whole genome amplification(WGA) was performed using a Sureplex DNA Amplification System and analyzed by PCR, targeting the deletion of the exons 5 and 6 in the GHR gene as well as PGT by Next-generation Sequencing(Illumina). Eleven embryos were collected and analyzed. 27.3% were the wild type for GHR, 45.5%were heterozygotes, and 18.2% homozygous mutants. One embryo yielded no results. Three 2-embryos transfers were performed; 2 normal homozygous and four heterozygous carriers were selected for transfer. The first two transfers were unsuccessful, whereas the final transfer with two heterozygous embryos resulted in clinical pregnancy. The genomic composition of the fetus was verified,applying the same techniques using amniocytes, extracted after 21 wk of the ongoing pregnancy. The fetus was confirmed as GHR deletion in exon 5-6,carrier. A non-affected baby was born.CONCLUSION Here, we present a case demonstrating that using WGA as a template in addition to PCR targeting specific gene regions, exons 5 and 6 on the GHR gene, could identify LS carrier embryos. This provides evidence that WGA and PCR serve as an excellent tool to detect this specific monogenic disease in IVF embryos, th

关 键 词：GROWTH HORMONE INSENSITIVITY GROWTH HORMONE receptor mutations Intragenic DELETIONS Molecular DIAGNOSIS EMBRYO DIAGNOSIS Laron syndrome Case report 

分 类 号：R73[医药卫生—肿瘤]