Analysis of cosegregation of intragenic DNA sequence variations as markers of maternal cell contamination in prenatal diagnosis of β-thalassemia

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Abstract

Prenatal diagnosis of 3 HBB gene mutations causing β-thalassemia and hemoglobin D Punjab segregated in a South Indian nuclear family is reported along with a method identified as control for maternal cell contamination (MCC). Amplicons of the HBB gene from genomic DNA obtained from the blood of a thalassemic first child (proband), both parents, and a chorionic villus sample of their second pregnancy were directly sequenced. A test for MCC was performed by genotyping polymorphic microsatellite markers (D21S11 and D21S1270) by quantitative fluorescence polymerase chain reaction (QF-PCR) and capillary gel electrophoresis. The pedigree analysis showed proband as a compound heterozygote of NG-000007.3:g.70691G>C and NG-000007.3:g.72128T>C mutations; showed the father as a compound heterozygote of NG-000007.3:g.72128T>C and NG-000007.3:g.71938G>C mutations; and showed the mother as a heterozygous carrier of the NG-000007.3:g.70691G>C mutation. The fetus inherited a normal maternal allele and a mutant paternal allele NG-000007.3:g.72128T>C and was ascertained a carrier of β-thalassemia. Analysis of cosegregation of 5 other single nucleotide polymorphisms (SNPs) in the family, including NG-000007.3:g.70603T>C, NG-000007.3:g.71055G>C, NG-000007.3:g.71113T>G, NG-000007.3:g.72332G>A, and NG-000007.3:g.72334A>C, defined the disease allele haplotypes. QF-PCR showed no extra maternal alleles in the fetal sample. Prenatal diagnosis of mutations and an absence of MCC was confirmed by cosegregation of the SNPs, suggesting the utility of a panel of such polymorphisms that can serve to identify MCC quickly and reliably.

Original languageEnglish
Pages (from-to)150-155
Number of pages6
JournalTranslational Research
Volume157
Issue number3
DOIs
Publication statusPublished - 03-2011

Fingerprint

Thalassemia
DNA sequences
Prenatal Diagnosis
Contamination
Polymorphism
Mothers
Polymerase chain reaction
Mutation
Alleles
Nucleotides
Genes
Fluorescence
Heterozygote
Single Nucleotide Polymorphism
Electrophoresis
Microsatellite Repeats
Blood
Gels
Chorionic Villi
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Medicine(all)
  • Biochemistry, medical
  • Public Health, Environmental and Occupational Health

Cite this

@article{d8787f4287b2460599473dcc1db0dea2,
title = "Analysis of cosegregation of intragenic DNA sequence variations as markers of maternal cell contamination in prenatal diagnosis of β-thalassemia",
abstract = "Prenatal diagnosis of 3 HBB gene mutations causing β-thalassemia and hemoglobin D Punjab segregated in a South Indian nuclear family is reported along with a method identified as control for maternal cell contamination (MCC). Amplicons of the HBB gene from genomic DNA obtained from the blood of a thalassemic first child (proband), both parents, and a chorionic villus sample of their second pregnancy were directly sequenced. A test for MCC was performed by genotyping polymorphic microsatellite markers (D21S11 and D21S1270) by quantitative fluorescence polymerase chain reaction (QF-PCR) and capillary gel electrophoresis. The pedigree analysis showed proband as a compound heterozygote of NG-000007.3:g.70691G>C and NG-000007.3:g.72128T>C mutations; showed the father as a compound heterozygote of NG-000007.3:g.72128T>C and NG-000007.3:g.71938G>C mutations; and showed the mother as a heterozygous carrier of the NG-000007.3:g.70691G>C mutation. The fetus inherited a normal maternal allele and a mutant paternal allele NG-000007.3:g.72128T>C and was ascertained a carrier of β-thalassemia. Analysis of cosegregation of 5 other single nucleotide polymorphisms (SNPs) in the family, including NG-000007.3:g.70603T>C, NG-000007.3:g.71055G>C, NG-000007.3:g.71113T>G, NG-000007.3:g.72332G>A, and NG-000007.3:g.72334A>C, defined the disease allele haplotypes. QF-PCR showed no extra maternal alleles in the fetal sample. Prenatal diagnosis of mutations and an absence of MCC was confirmed by cosegregation of the SNPs, suggesting the utility of a panel of such polymorphisms that can serve to identify MCC quickly and reliably.",
author = "Saadi, {Abdul V.} and Girisha, {Katta M.} and Gopinath, {Puthiya M.} and Kapaettu Satyamoorthy",
year = "2011",
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doi = "10.1016/j.trsl.2010.12.005",
language = "English",
volume = "157",
pages = "150--155",
journal = "Translational Research",
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T1 - Analysis of cosegregation of intragenic DNA sequence variations as markers of maternal cell contamination in prenatal diagnosis of β-thalassemia

AU - Saadi, Abdul V.

AU - Girisha, Katta M.

AU - Gopinath, Puthiya M.

AU - Satyamoorthy, Kapaettu

PY - 2011/3

Y1 - 2011/3

N2 - Prenatal diagnosis of 3 HBB gene mutations causing β-thalassemia and hemoglobin D Punjab segregated in a South Indian nuclear family is reported along with a method identified as control for maternal cell contamination (MCC). Amplicons of the HBB gene from genomic DNA obtained from the blood of a thalassemic first child (proband), both parents, and a chorionic villus sample of their second pregnancy were directly sequenced. A test for MCC was performed by genotyping polymorphic microsatellite markers (D21S11 and D21S1270) by quantitative fluorescence polymerase chain reaction (QF-PCR) and capillary gel electrophoresis. The pedigree analysis showed proband as a compound heterozygote of NG-000007.3:g.70691G>C and NG-000007.3:g.72128T>C mutations; showed the father as a compound heterozygote of NG-000007.3:g.72128T>C and NG-000007.3:g.71938G>C mutations; and showed the mother as a heterozygous carrier of the NG-000007.3:g.70691G>C mutation. The fetus inherited a normal maternal allele and a mutant paternal allele NG-000007.3:g.72128T>C and was ascertained a carrier of β-thalassemia. Analysis of cosegregation of 5 other single nucleotide polymorphisms (SNPs) in the family, including NG-000007.3:g.70603T>C, NG-000007.3:g.71055G>C, NG-000007.3:g.71113T>G, NG-000007.3:g.72332G>A, and NG-000007.3:g.72334A>C, defined the disease allele haplotypes. QF-PCR showed no extra maternal alleles in the fetal sample. Prenatal diagnosis of mutations and an absence of MCC was confirmed by cosegregation of the SNPs, suggesting the utility of a panel of such polymorphisms that can serve to identify MCC quickly and reliably.

AB - Prenatal diagnosis of 3 HBB gene mutations causing β-thalassemia and hemoglobin D Punjab segregated in a South Indian nuclear family is reported along with a method identified as control for maternal cell contamination (MCC). Amplicons of the HBB gene from genomic DNA obtained from the blood of a thalassemic first child (proband), both parents, and a chorionic villus sample of their second pregnancy were directly sequenced. A test for MCC was performed by genotyping polymorphic microsatellite markers (D21S11 and D21S1270) by quantitative fluorescence polymerase chain reaction (QF-PCR) and capillary gel electrophoresis. The pedigree analysis showed proband as a compound heterozygote of NG-000007.3:g.70691G>C and NG-000007.3:g.72128T>C mutations; showed the father as a compound heterozygote of NG-000007.3:g.72128T>C and NG-000007.3:g.71938G>C mutations; and showed the mother as a heterozygous carrier of the NG-000007.3:g.70691G>C mutation. The fetus inherited a normal maternal allele and a mutant paternal allele NG-000007.3:g.72128T>C and was ascertained a carrier of β-thalassemia. Analysis of cosegregation of 5 other single nucleotide polymorphisms (SNPs) in the family, including NG-000007.3:g.70603T>C, NG-000007.3:g.71055G>C, NG-000007.3:g.71113T>G, NG-000007.3:g.72332G>A, and NG-000007.3:g.72334A>C, defined the disease allele haplotypes. QF-PCR showed no extra maternal alleles in the fetal sample. Prenatal diagnosis of mutations and an absence of MCC was confirmed by cosegregation of the SNPs, suggesting the utility of a panel of such polymorphisms that can serve to identify MCC quickly and reliably.

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