Evaluation of single-nucleotide polymorphisms as internal controls in prenatal diagnosis of fetal blood groups.
Transfusion
; 53(2): 353-62, 2013 Feb.
Article
en En
| MEDLINE
| ID: mdl-22691192
ABSTRACT
BACKGROUND:
Determination of fetal blood groups in maternal plasma samples critically depends on adequate amplification of fetal DNA. We evaluated the routine inclusion of 52 single-nucleotide polymorphisms (SNPs) as internal reference in our polymerase chain reaction (PCR) settings to obtain a positive internal control for fetal DNA. STUDY DESIGN ANDMETHODS:
DNA from 223 plasma samples of pregnant women was screened for RHD Exons 3, 4, 5, and 7 in a multiplex PCR including 52 SNPs divided into four primer pools. Amplicons were analyzed by single-base extension and the GeneScan method in a genetic analyzer. Results of D screening were compared to standard RHD genotyping of amniotic fluid or real-time PCR of fetal DNA from maternal plasma.RESULTS:
The vast majority of all samples (97.8%) demonstrated differences in maternal and fetal SNP patterns when tested with four primer pools. These differences were not observed in less than 2.2% of the samples most probably due to an extraction failure for adequate amounts of fetal DNA. Comparison of the fetal genotypes with independent results did not reveal a single false-negative case among samples (n = 42) with positive internal control and negative fetal RHD typing.CONCLUSION:
Coamplification of 52 SNPs with RHD-specific sequences for fetal blood group determination introduces a valid positive control for the amplification of fetal DNA to avoid false-negative results. This new approach does not require a paternal blood sample. It may also be applicable to other assays for fetal genotyping in maternal blood samples.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Diagnóstico Prenatal
/
Sistema del Grupo Sanguíneo Rh-Hr
/
Antígenos de Grupos Sanguíneos
/
Polimorfismo de Nucleótido Simple
/
Sangre Fetal
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
Límite:
Female
/
Humans
/
Pregnancy
Idioma:
En
Revista:
Transfusion
Año:
2013
Tipo del documento:
Article
País de afiliación:
Alemania