MATERIALS AND METHODS
a. Study population: This study was carried out on 40 primigravida pregnant females who attended the Antenatal Clinic at Elshatby Maternity University Hospital, Alexandria. The age of subjects was above 18 years and below 35 years. All subjects had missed abortion from the 7th week + 1day to the 13th week + 6 days with a history of positive fetal cardiac activity that had stopped suddenly.
b. Methods: All the patients in the study were subjected to the following: detailed history taking, thorough clinical examination, ultrasound for fetal viability and gestational age, and finally, blood samples were collected from all pregnant women enrolled in the study for DNA extraction, and genotype analysis. FVL and MTHFR gene mutations were assayed based on polymerase chain reaction (PCR) and reverse-hybridization. The frequency of homozygous and heterozygous gene mutations, as well as, the co-expression of mutations was determined. The procedure includes three steps: (1) DNA isolation, (2) PCR amplification using biotinylated primers, (3) hybridization of amplification products to a test strip containing allele-specific oligonucleotide probes immobilized as an array of parallel lines. The assays covered two mutations: FV G1691A (Factor V Leiden Kit Roche®), and MTHFR C677T (DiaplexQ™ MTHFR genotyping Kit®). The kit for FV-PTH-MTHFR Strip Assay was purchased from ViennaLab Diagnostics GmbH Gaudenzdorfer Guertel 43-45 A-1120 Vienna, Austria.
c. Statistical analysis of the data: Data were fed to the computer and analyzed using IBM SPSS software package version 20.0. (Armonk, NY: IBM Corp)(11) Qualitative data were described using the number and percent. Quantitative data were described using range (minimum and maximum), mean, standard deviation, median, and interquartile range (IQR).