title: Non-invasive Prenatal Diagnosis of Feto-Maternal Platelet Incompatibility by Cold High Resolution Melting Analysis.
abstract: Fetal and Neonatal alloimmune thrombocytopenia (FNAIT) is a condition which could occur when pregnant women develop an alloimmunization against paternally inherited antigens of the fetal platelets. Approximately 80 % of FNAIT cases are caused by anti-HPA-1a, about 15 % by anti-HPA-5b and 5 % by other HPA antibodies. Only 2 % of the total population is HPA-1a negative (HPA-1b1b). The HPA-1a allele differs by one single nucleotide from HPA-1b allele, yet it represents around 27 % of total severe thrombocytopenias. HPA-1 was studied in serum cDNA from 12 volunteer pregnant women to determine their HPA-1 genotype by HRM (high resolution melting) PCR. When an homozygous HPA-1 gene was detected in a mother, a COLD HRM was performed to determine whether or not the fetal genotype differs from the mother's.The differences in the melting curve shapes allow us to accurately distinguish the three pregnants genotypes. The fetal heterozygous genotype of homozygous pregnant women was correctly detected by COLD PCR HRM in maternal serum. HPA-1 genotyping by HRM may be a useful aproach for genotyping all pregnant women in inexpensively. Moreover, when HPA-1 homozygosis was detected in a pregnant woman, fetal heterozygosis may be diagnosed by COLD HRM to select pregnancies for preventive monitoring.
authors.affiliation: Department of Clinical Biochemistry, Instituto de Biomedicina de Sevilla (IBiS/Csic/SAS/University of Seville), University Hospital Virgen del Rocío of Seville, Seville, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Ferro
authors.affiliation: Department of Clinical Biochemistry, Instituto de Biomedicina de Sevilla (IBiS/Csic/SAS/University of Seville), University Hospital Virgen del Rocío of Seville, Seville, Spain. hadacmacher@icloud.com.
authors.forename: Hada C
authors.initials: HC
authors.lastname: Macher
authors.affiliation: Immunohematology Unit of University Hospital Virgen del Rocío of Seville, Seville, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Noguerol
authors.affiliation: Department of Clinical Biochemistry, Instituto de Biomedicina de Sevilla (IBiS/Csic/SAS/University of Seville), University Hospital Virgen del Rocío of Seville, Seville, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Jimenez-Arriscado
authors.affiliation: Department of Clinical Biochemistry, Instituto de Biomedicina de Sevilla (IBiS/Csic/SAS/University of Seville), University Hospital Virgen del Rocío of Seville, Seville, Spain.
authors.forename: Patrocinio
authors.initials: P
authors.lastname: Molinero
authors.affiliation: Department of Clinical Biochemistry, Instituto de Biomedicina de Sevilla (IBiS/Csic/SAS/University of Seville), University Hospital Virgen del Rocío of Seville, Seville, Spain.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Guerrero
authors.affiliation: Department of Clinical Biochemistry, Instituto de Biomedicina de Sevilla (IBiS/Csic/SAS/University of Seville), University Hospital Virgen del Rocío of Seville, Seville, Spain.
authors.forename: Amalia
authors.initials: A
authors.lastname: Rubio
mesh_terms: D016824:Antigens, Human Platelet / Q000097:blood / Q000235:genetics / Q000276:immunology; D001792:Blood Platelets / Q000378:metabolism*; D003362:Cost-Benefit Analysis; D004247:DNA / Q000097:blood / Q000235:genetics; D005260:Female; D005838:Genotype; D060005:Genotyping Techniques / Q000191:economics / Q000379:methods; D006801:Humans; D007231:Infant, Newborn; D039661:Integrin beta3; D008431:Maternal-Fetal Exchange / Q000235:genetics / Q000276:immunology*; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods; D054098:Thrombocytopenia, Neonatal Alloimmune / Q000097:blood / Q000235:genetics / Q000276:immunology*
chemical_list: D016824:Antigens, Human Platelet; C061248:ITGB3 protein, human; D039661:Integrin beta3; D004247:DNA
keywords: Alloimmune thrombocytopenia; Fetal circulating DNA