Abstract |
Our purpose was to develop and evaluate isolation and enrichment of fetal erythroblasts and a nested polymerase chain reaction (PCR) approach using fetal erythroblasts for detecting the beta-globin gene mutations for a noninvasive prenatal diagnosis of hemoglobinopathies. Maternal blood at different periods of gestation was layered on a Percoll density gradient for enrichment of fetal nucleated RBCs (NRBCs). A combination of 3 monoclonal antibodies (CD45- peridinin chlorophyll protein, glycophorin A- phycoerythrin, and anti- hemoglobin F- fluorescein isothiocyanate) was used for flow cytometric sorting of fetal NRBCs from enriched cells. Different nested PCR-based approaches were used for identification of fetal mutations. Owing to heterogeneity of beta-thalassemia mutations in the population of India, we had to screen for 12 mutations and were able to give an accurate diagnosis in 84 (84.0%) of 100 cases when compared with chorionic villus sampling or cordocentesis and DNA analysis.This nested PCR approach enabled amplification of small quantities of DNA from fetal erythroblasts, providing a cost-effective method for noninvasive diagnosis.
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Authors | Edna D'Souza, Pratibha M Sawant, Anita H Nadkarni, Ajit Gorakshakar, Dipika Mohanty, Kanjaksha Ghosh, Roshan B Colah |
Journal | American journal of clinical pathology
(Am J Clin Pathol)
Vol. 130
Issue 2
Pg. 202-9
(Aug 2008)
ISSN: 0002-9173 [Print] England |
PMID | 18628088
(Publication Type: Comparative Study, Evaluation Study, Journal Article)
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Chemical References |
- Antibodies, Monoclonal
- Globins
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Topics |
- Antibodies, Monoclonal
- Cost-Benefit Analysis
- Erythroblasts
- Female
- Fetal Blood
- Genetic Testing
- Globins
(genetics)
- Hemoglobinopathies
(diagnosis)
- Humans
- India
- Minisatellite Repeats
- Mutation
- Polymerase Chain Reaction
- Pregnancy
- Prenatal Diagnosis
(economics, methods)
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