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Reliable Detection of Paternal SNPs within Deletion Breakpoints for Non-Invasive Prenatal Exclusion of Homozygous α0-Thalassemia in Maternal Plasma
- Source :
- PLoS ONE
- Publication Year :
- 2011
- Publisher :
- Public Library of Science, 2011.
-
Abstract
- Reliable detection of large deletions from cell-free fetal DNA (cffDNA) in maternal plasma is challenging, especially when both parents have the same deletion owing to a lack of specific markers for fetal genotyping. In order to evaluate the efficacy of a non-invasive prenatal diagnosis (NIPD) test to exclude α-thalassemia major that uses SNPs linked to the normal paternal α-globin allele, we established a novel protocol to reliably detect paternal SNPs within the (--(SEA)) breakpoints and performed evaluation of the diagnostic potential of the protocol in a total of 67 pregnancies, in whom plasma samples were collected prior to invasive obstetrics procedures in southern China. A group of nine SNPs identified within the deletion breakpoints were scanned to select the informative SNPs in each of the 67 couples DNA by multiplex PCR based mini-sequencing technique. The paternally inherited SNP allele from cffDNA was detected by allele specific real-time PCR. A protocol for reliable detection of paternal SNPs within the (--(SEA)) breakpoints was established and evaluation of the diagnostic potential of the protocol was performed in a total of 67 pregnancies. In 97% of the couples one or more different SNPs within the deletion breakpoint occurred between paternal and maternal alleles. Homozygosity for the (--(SEA)) deletion was accurately excluded in 33 out of 67 (49.3%, 95% CI, 25.4-78.6%) pregnancies through the implementation of the protocol. Protocol was completely concordant with the traditional reference methods, except for two cases that exhibited uncertain results due to sample hemolysis. This method could be used as a routine NIPD test to exclude gross fetal deletions in α-thalassemia major, and could further be employed to test for other diseases due to gene deletion.
- Subjects :
- Adult
Male
China
Heterozygote
Gene Expression
Mothers
Single-nucleotide polymorphism
Prenatal diagnosis
Alpha-thalassemia
Biology
Bioinformatics
Polymerase Chain Reaction
Polymorphism, Single Nucleotide
Sensitivity and Specificity
Plasma
Autosomal Recessive
Gene Frequency
alpha-Thalassemia
Pregnancy
Nucleic Acids
Prenatal Diagnosis
Multiplex polymerase chain reaction
Molecular Cell Biology
medicine
Humans
Genetic Testing
Allele
Genotyping
Allele frequency
Alleles
Genetics
Clinical Genetics
Multidisciplinary
Nucleotides
DNA
Hematology
medicine.disease
Hematopoiesis
Cell-free fetal DNA
Medicine
Female
Gene Deletion
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 6
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....dbe7868ad9e088f3550c2796a28f4a4a