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).