title: Evaluation of fetal hypermethylated RASSF1A in pre-eclampsia and its relationship with placental protein-13, pregnancy associated plasma protein-A and urine protein.
abstract: OBJECTIVE
  Cell free fetal DNA (cffDNA) and its hypermethylated RASSF1A gene signify a recent advancement in non-invasive prenatal diagnosis of feto-placental anomalies like pre-eclampsia. The study uses hypermethylated RASSF1A gene to quantify cffDNA and to assess its relationship with placental and urine proteins in pre-eclampsia cases.
  
  
  METHODS
  DNA was isolated from plasma samples of clinically diagnosed cases of pre-eclampsia (n=103) and normal pregnancy (n=616) from 21weeks of gestation. Through methylation sensitive enzyme (BstUI) digestion; followed by real time-polymerase chain reaction (RT-PCR), quantification of hypermethylated RASSF1A was done. Immunoassays determined: placental protein-13 (pp-13) and pregnancy associated plasma protein A (PAPP-A) and pyrogallol red molybdate assay for 24h urine protein.
  
  
  RESULTS
  Highly significant differences between control and pre-eclampsia cases for hypermethylated RASSF1A concentrations were found; Group I: 33±7.35 vs 74.46±16.71, Group II: 53.75±16.65 vs 244.22±35.68, Group III: 93.25±19.08 vs 412.31±80.18, Group IV: 144.30±18.13 vs 1056.89±153.78, Group V: 307.55±40.76 vs 2763.76±259.76copies/ml. Multivariate Pearson's correlation analysis of hypermethylated RASSF1A with pp-13, PAPP-A and urine proteins showed positive and very highly significant (P<0.001) associations.
  
  
  CONCLUSIONS
  Diagnostic potential of fetal specific, hypermethylated RASSF1A was evaluated. Its positive relationship with placental and urine proteins submit the case for considering it as a reliable marker for pre-eclampsia.
authors.affiliation: Base Hospital, Delhi Cantt, School of Medicine & Paramedical Health Sciences, Guru Gobind Singh Indraprastha University, Delhi, India. Electronic address: manuseema26@yahoo.com.
authors.forename: Seema
authors.initials: S
authors.lastname: Saraswathy
authors.affiliation: Department of Pathology, Armed Forces Medical College (AFMC), Pune, India.
authors.forename: Kavita
authors.initials: K
authors.lastname: Sahai
authors.affiliation: Department of Paediatrics, PGIMER, RML Hospital, Delhi, India.
authors.forename: Tribhuvan Pal
authors.initials: TP
authors.lastname: Yadav
authors.affiliation: Dept of Obstetrics and Gynaecology, Base Hospital and Army College of Medical Sciences (ACMS), Delhi Cantt, India.
authors.forename: Devendra
authors.initials: D
authors.lastname: Arora
authors.affiliation: North DMC Medical College, Hindu Rao Hospital, Delhi, India.
authors.forename: Suman Lata
authors.initials: SL
authors.lastname: Mendiratta
authors.affiliation: Molecular Diagnostics, Genetix Biotech Asia, New Delhi, India.
authors.forename: Samar Husain
authors.initials: SH
authors.lastname: Naqvi
authors.affiliation: Department of Biosciences, Jamia Millia Islamia, Delhi, India.
authors.forename: Shilpie
authors.initials: S
authors.lastname: Biswas
authors.affiliation: Institute of Nuclear Medicine & Allied Sciences (INMAS), Defence Research & Development Organization (DRDO), Ministry of Defence, Govt of India, Delhi, India.
authors.forename: Manu
authors.initials: M
authors.lastname: Krishnan
authors.affiliation: Department of Zoology, Mar Thoma College, Tiruvalla, Kerala, India.
authors.forename: Kurian Mathew
authors.initials: KM
authors.lastname: Abraham
mesh_terms: D000328:Adult; D016022:Case-Control Studies; D019175:DNA Methylation; D005260:Female; D005333:Fetus / Q000378:metabolism; D037161:Galectins / Q000097:blood; D006801:Humans; D011225:Pre-Eclampsia / Q000097:blood* / Q000175:diagnosis*; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000097:blood; D011266:Pregnancy-Associated Plasma Protein-A / Q000378:metabolism; D011296:Prenatal Diagnosis / Q000379:methods*; D011507:Proteinuria / Q000652:urine; D025521:Tumor Suppressor Proteins / Q000235:genetics*; D055815:Young Adult
chemical_list: D037161:Galectins; C038195:LGALS13 protein, human; D011257:Pregnancy Proteins; C411762:RASSF1 protein, human; D025521:Tumor Suppressor Proteins; D011266:Pregnancy-Associated Plasma Protein-A
keywords: Cell free fetal DNA; Hypermethylated RASSF1A; Placental proteins; Pre-eclampsia