الفهرس 
Repeated Spontaneous MiscarriageOnly 14 pages are availabe for public view
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Abstract
Miscarriage is one of the leading problems in women’s health issues, 9-13% of women in the reproductive age group experience one clinically recognized loss, 5% experience two or more losses, and 1 to 2% suffer three or more losses . Although several medical causes have been established, up to 50% of cases of repeated spontaneous miscarriages (RSM) still remain unexplained after standard gynecological, hormonal, and karyotypic investigations.
Thrombophilia is a multifactorial disease that involves environmental and genetic risk factors, the incidence of thrombosis in individuals having genetic defects is highly variable. This depends on the particular genotype, coexistence of other genetic defects, and influence of environmental risk factors such as oral contraceptives, trauma, surgery, and pregnancy.
The most frequent reasons of congenital thrombophilia are factor V Leiden (G1691A), prothrombin gene mutation (G20210A), and 5, 10-methylenetetrahydrofolate reductase gene mutation (C677T).
The aim of our study is to clarify the value of the aetiological diagnosis of mutations in thrombophilic factors genes [factor V Leiden (FVL) gene, the prothrombin (PTH) gene and methylenetetrahydrofolate reductase (MTHFR) gene] in women with unexplained first trimester repeated spontaneous miscarriage (RSM).
Our study included 40 women, classified as normal control group who had history of normal pregnancy (no pregnancy disorders) (n=20) and cases of unexplained first trimester repeated spontaneous miscarriage (n=20).
Our study excluded women with previous history of thrombosis, pregnancy at the time of investigation, using oral contraceptives or with any known cause of miscarriage.
Blood samples (5 ml) were taken; of which 3 ml were put on sodium citrate for determination of PT, PC and APTT. However, the other 2 ml were put on EDTA for mutation detection of factor V (FV), prothrombin (PTH) and methelynetetrahydrofolate reductase (MTHFR) genes based on the reverse- hybridization principle using (HVD strip kit). The mutation detection assay covered the 3 mutations: FV (G1691A), PTH (G20210) and MTHFR (C677T) and included five steps:
DNA isolation (using 100 µl blood sample for extraction), PCR amplification (in a single multiplex reaction using biotinylated primers) and hybridization of amplified products to a test strip containing allele-specific oligonucleotide probes immobilized as an array of parallel lines, bound biotinylated sequences were detected using streptavidin-alkaline phosphatase and color substrates and interpretation of results.
Our data showed significant increase of allele frequency as well as homozygosity and heterozygosity of FV, PTH gene and MTHFR mutations in women with unexplained (RSM) as compared to normal control group.
In the case of multiple genes mutation, our results showed significantly increased prevalence of multiple thrombophilic genes mutation (of FVL, PTH and MTHFR C677T) in women with RSM compared to normal control group.
Conclusion:
Our results showed increased allele frequencies of thrombophilic mutation of factor V, prothrombin and methylenetetrahydrofolate reductase genes, in addition to increased genotype frequencies (both homozygote and heterozygote). Also our results showed increased frequency of multiple genes mutation among women with unexplained first trimester repeated spontaneous miscarriage (RSM).
These results emphasizes the role of thrombophilia as a cause of RSM which is still a common complication of pregnancy, in order to establish a possible protocol that supports the aetiological diagnosis of RSM associated with thromboprophylaxis of these patients.
Recommendations:
• Studying the effects of thrombophilia treatment on adverse pregnancy outcome.
• Correlative study of congenital thrombophilic factors in the three trimesters of pregnancy.
• The relative value of a thrombophilia screening programme to other healthcare programmes needs to be established.