title: APCR, factor V gene known and novel SNPs and adverse pregnancy outcomes in an Irish cohort of pregnant women.
abstract: BACKGROUND
  Activated Protein C Resistance (APCR), a poor anticoagulant response of APC in haemostasis, is the commonest heritable thrombophilia. Adverse outcomes during pregnancy have been linked to APCR. This study determined the frequency of APCR, factor V gene known and novel SNPs and adverse outcomes in a group of pregnant women.
  
  
  METHODS
  Blood samples collected from 907 pregnant women were tested using the Coatest Classic and Modified functional haematological tests to establish the frequency of APCR. PCR-Restriction Enzyme Analysis (PCR-REA), PCR-DNA probe hybridisation analysis and DNA sequencing were used for molecular screening of known mutations in the factor V gene in subjects determined to have APCR based on the Coatest Classic and/or Modified functional haematological tests. Glycosylase Mediated Polymorphism Detection (GMPD), a SNP screening technique and DNA sequencing, were used to identify SNPs in the factor V gene of 5 APCR subjects.
  
  
  RESULTS
  Sixteen percent of the study group had an APCR phenotype. Factor V Leiden (FVL), FV Cambridge, and haplotype (H) R2 alleles were identified in this group. Thirty-three SNPs; 9 silent SNPs and 24 missense SNPs, of which 20 SNPs were novel, were identified in the 5 APCR subjects. Adverse pregnancy outcomes were found at a frequency of 35% in the group with APCR based on Classic Coatest test only and at 45% in the group with APCR based on the Modified Coatest test. Forty-eight percent of subjects with FVL had adverse outcomes while in the group of subjects with no FVL, adverse outcomes occurred at a frequency of 37%.
  
  
  CONCLUSIONS
  Known mutations and novel SNPs in the factor V gene were identified in the study cohort determined to have APCR in pregnancy. Further studies are required to investigate the contribution of these novel SNPs to the APCR phenotype. Adverse outcomes including early pregnancy loss (EPL), preeclampsia (PET) and intrauterine growth restriction (IGUR) were not significantly more frequent in subjects with APCR compared to normal pregnant women however Pregnancy induced hypertension (PIH) was found to be associated with FVL in our study group.
authors.affiliation: Molecular Diagnostics Research Group, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland. sara.sedano@nuigalway.ie
authors.forename: Sara
authors.initials: S
authors.lastname: Sedano-Balbás
authors.forename: Mark
authors.initials: M
authors.lastname: Lyons
authors.forename: Brendan
authors.initials: B
authors.lastname: Cleary
authors.forename: Margaret
authors.initials: M
authors.lastname: Murray
authors.forename: Geraldine
authors.initials: G
authors.lastname: Gaffney
authors.forename: Majella
authors.initials: M
authors.lastname: Maher
mesh_terms: D000022:Abortion, Spontaneous / Q000453:epidemiology / Q000235:genetics; D020016:Activated Protein C Resistance / Q000097:blood / Q000150:complications / Q000453:epidemiology / Q000235:genetics*; D016022:Case-Control Studies; D016009:Chi-Square Distribution; D004252:DNA Mutational Analysis; D005165:Factor V / Q000235:genetics*; D005260:Female; D005317:Fetal Growth Retardation / Q000453:epidemiology / Q000235:genetics; D005820:Genetic Testing; D006239:Haplotypes; D006801:Humans; D007494:Ireland / Q000453:epidemiology; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011225:Pre-Eclampsia / Q000453:epidemiology / Q000235:genetics; D011247:Pregnancy; D011250:Pregnancy Complications, Hematologic / Q000097:blood / Q000453:epidemiology / Q000235:genetics*; D011256:Pregnancy Outcome / Q000453:epidemiology / Q000235:genetics; D000091402:Prohibitins; D018709:Statistics, Nonparametric
chemical_list: C000717331:PHB2 protein, human; D000091402:Prohibitins; C119012:factor V Cambridge; C095381:factor V Leiden; D005165:Factor V