RESUMEN
BACKGROUND: Fetal human platelet antigen (HPA) genotyping is required to determine whether the fetus is at risk and whether prenatal interventions to prevent fetal bleeding are required in pregnant women with a history of fetal and neonatal alloimmune thrombocytopenia (FNAIT). Methods for noninvasive genotyping of HPA alleles with the use of maternal plasma cell-free DNA were published recently but do lack internal controls to exclude false-negative results. STUDY DESIGN AND METHODS: Cell-free DNA was isolated from plasma of four pregnant women with a history of FNAIT caused by anti-HPA-1a and controls. A primer panel was designed to target sequences flanking single-nucleotide polymorphisms (SNPs)/exonic regions of ITGB3 (HPA-1), ITGA2B (HPA-3), ITGA2 (HPA-5), CD109 (HPA-15), RHD, RHCE, KEL, DARC, SLC14A1, GYPA, GYPB, and SRY. These regions and eight anonymous SNPs were massively parallel sequenced by semiconductor technology. RESULTS: The mean (±SD) number of reads for targeted SNPs was 5255 (±2838). Fetal DNA was detected at a median of 4.5 (range, 2-8) polymorphic loci. The mean fractional fetal DNA concentration in cell-free maternal plasma was 8.36% (range, 4.79%-15.9%). For HPA-1, nonmaternal ITGB3 sequences (c.176T, HPA-1a) were detected in all HPA-1ab fetuses. One HPA-1bb fetus was unequivocally identified, showing the pregnancy was not at risk of FNAIT. CONCLUSION: We have successfully established massively parallel sequencing as a novel reliable method for noninvasive genotyping of fetal HPA-1a alleles. This technique may also allow the safe detection of other fetal blood group polymorphisms frequently involved in FNAIT and hemolytic disease of the newborn.
Asunto(s)
Antígenos de Plaqueta Humana/genética , Enfermedades Fetales/diagnóstico , Técnicas de Genotipaje/métodos , Secuenciación de Nucleótidos de Alto Rendimiento , Diagnóstico Prenatal/métodos , Análisis de Secuencia de ADN/métodos , Trombocitopenia Neonatal Aloinmune/diagnóstico , Análisis Químico de la Sangre/métodos , Estudios de Casos y Controles , Sistema Libre de Células , Femenino , Enfermedades Fetales/genética , Genotipo , Humanos , Recién Nacido , Integrina beta3 , Polimorfismo de Nucleótido Simple , Embarazo/sangre , Reproducibilidad de los Resultados , Trombocitopenia Neonatal Aloinmune/genéticaRESUMEN
Endometriosis is the second most frequent benign disease of the female of reproductive age. In spite of extensive research, the etiology of this disease is still largely enigmatic. A causal therapy has not been found yet. Several drug therapy approaches are available but none offers a permanent cure of endometriosis. In addition, the range of side effects of the particular drug therapies has to be considered individually. The treatment of patients with endometriosis cannot be standardized but has to differentiate the individual factors and life circumstances of the patients.
