Potential biomarker of circulating hsa-miR-1273g-3p level for detection of recurrent epithelial ovarian cancer.

PURPOSE: Ovarian cancer (OC) is first gynaecologic cancer that causes women death and epithelial ovarian cancer (EOC) is the most lethal ovarian cancer type. While treatment is commonly successful, some cases (10-20%) show resistance to chemotherapy which is followed by recurrence. MicroRNA (miRNA) based diagnosis methods are slightly important for recurrent ovarian cancer diagnosis. We aimed to detect novel circulating miRNAs to be used as an early diagnosis and prediction tools for recurrent EOC. METHODS: In this study, recurrent EOC serum samples and healthy control serum samples were compared for miRNA expression analysis by microarray. Microarray results were analyzed by bioinformatics tools and differentially expressed hsa-miR-1273g-3p was obtained. After microarray analysis, differentially expressed hsa-miR-1273g-3p was validated by Real-Time PCR (RT-qPCR). The relation between target genes of hsa-miR-1273g-3p and ovarian cancer were examined by Pathway Studio® (v.11.4.0.8). RESULTS: The expression of hsa-miR-1273g-3p was found to be significantly down-regulated by t test Bonferroni FWER corrected p < 0.05 and fold change > 2, in recurrence EOC compare with healthy controls groups. The RT-qPCR results confirmed that relative expressions of the serum hsa-miR-1273g-3p were significantly down-regulated in patients with recurrent EOC (p = 0.0275). Serum hsa-miR-1273g-3p levels could discriminate patients with recurrent EOC from healthy controls, with a power area under the curve (AUC) of 0.7. CONCLUSION: This study suggested that hsa-miR-1273g-3p plays a significant role in regulation of related genes, which are TNF-alfa, COL1A1, MMP-2, MMP-9, with recurrent EOC outcome. hsa-miR-1273g-3p may be used as a prognostic marker for recurrent EOC after chemotherapy.

Clinical evaluations of cell-free fetal DNA quantities in pre-eclamptic pregnancies.

AIM: Quantitative changes of cell-free fetal DNA in maternal plasma as an indicator for impending pre-eclampsia was reported in different studies. Cell-free fetal nucleic acids can be detected in maternal circulation during pregnancy. Our aim was to determine the higher rate of fetal DNA levels in maternal blood in pre-eclampsia compared to normal pregnancies and the clinical use of real-time polymerase chain reaction (PCR) in the Turkish population as a marker. MATERIAL AND METHODS: According to their gestational ages, the plasma levels of 30 pre-eclamptic women at 26-40 weeks of pregnancy were matched with 18 healthy pregnant women. Cell-free fetal DNA levels in maternal plasma were compared using real-time PCR technology. For the quantitative measurement of fetal DNA from maternal blood, the relative quantification PCR process was applied to all samples, using SRY and GAPDH genes. These patients were classified as pre-eclamptic and control groups and were matched according to weeks of pregnancy. RESULTS: Free fetal DNA levels of 30 pre-eclamptic patients were compared to healthy pregnant women and an average 3.06-fold increase was observed. During the second trimester, free fetal DNA levels were 1.5 times higher in pre-eclamptic patients. This increase was 3.5-fold during the third trimester. The DNA increase of pre-eclamptic patients was 4.1-fold and 3.4-fold during 29th-33rd and 34th-40th weeks, respectively. CONCLUSIONS: Cell-free fetal DNA in maternal blood could be used as a marker for identifying subjects at increased risk of developing pre-eclampsia.

Results of midtrimester emergency cerclage.

OBJECTIVE: To evaluate the outcomes of midtrimester emergency cerclage and to find out the contributing factors. STUDY DESIGN: Twenty-five patients presenting with cervical dilation and effacement with the membranes at the level of the external os or prolapsed into the vagina were included in the study. At the time of the cervical cerclage placement, gestational age ranged from 14-26 weeks. A good outcome was defined as the take-home baby rate, but also the interval between the time of the cervical cerclage placement and delivery and the gestational age at delivery were discussed. RESULTS: The mean gestational age at the time of the cervical cerclage placement was 21.2 +/- 2.73 weeks, the mean gestational age at delivery was 29.4 +/- 5.37 weeks, and the mean birth weight was 1,511 g. Eighteen patients had McDonald type cerclage, and 7 patients had both McDonald and Saling type cerclages. The overall take-home baby rate was 64%. Patients presenting with membranes at the level of external os had a take-home baby rate of 68.8%, whereas patients presenting with membranes prolapsed into the vagina had a take-home baby rate of 31.3%. CONCLUSION: Cervical dilation at midtrimester has a poor outcome which can be improved with emergency cerclage. Patients presenting with membranes prolapsed into the vagina have a decreased success rate. Total closure of the cervix may improve results.

Metabolic pathways of potential miRNA biomarkers derived from liquid biopsy in epithelial ovarian cancer.

Epithelial ovarian cancer (EOC) is the type of OC with the highest mortality rate. Due to the asymptomatic nature of the disease and few available diagnostic tests, it is mostly diagnosed at the advanced stage. Therefore, the present study aimed to discover predictive and/or early diagnostic novel circulating microRNAs (miRNAs or miRs) for EOC. Firstly, microarray analysis of miRNA expression levels was performed on 32 samples of female individuals: Eight plasma samples from patients with pathologically confirmed EOC (mean age, 45 (30-54) years), eight plasma samples from matched healthy individuals (HIs) (mean age, 44 (30-65) years), eight EOC tissue samples (mean age, 45 (30-54) years) and eight benign ovarian (mean age, 35 (17-70) years) neoplastic tissue samples A total of 31 significantly dysregulated miRNAs in serum and three miRNAs in tissue were identified by microarray. The results were validated using reverse transcription-quantitative PCR on samples from 10 patients with pathologically confirmed EOC (mean age, 47(30-54) years), 10 matched His (mean age, 40(26-65) years], 10 EOC tissue samples (mean age, 47(30-54) years) and 10 benign ovarian neoplastic tissue samples (mean age, 40(17-70) years). The 'Kyoto Encyclopedia of Genes and Genomes' (KEGG) database was used for target gene and pathway analysis. A total of three miRNAs from EOC serum (hsa-miR-1909-5p, hsa-miR-885-5p and hsa-let-7d-3p) and one microRNA from tissue samples (hsa-miR-200c-3p) were validated as significant to distinguish patients with EOC from HIs. KEGG pathway enrichment analysis showed seven significant pathways, which included 'prion diseases', 'proteoglycans in cancer', 'oxytocin signaling pathway', 'hippo signaling pathway', 'adrenergic signaling in cardiomyocytes', 'oocyte meiosis' and 'thyroid hormone signaling pathway', in which the validated miRNAs served a role. This supports the hypothesis that four validated miRNAs, have the potential to be a biomarker of EOC diagnosis and target for treatment.

Leriche's syndrome and twin pregnancy.

Leriche's syndrome is characterized by chronic obstruction of the abdominal aorta and iliac arteries. A patient with Leriche's syndrome presented with twin pregnancy and severe preeclampsia at 32 weeks' gestation. Cesarean delivery was performed and the patient was admitted to the intensive care unit. Magnetic resonance angiography showed total occlusion of the distal abdominal aorta, common, and external iliac arteries. There were extensive collateral vessels between the lumbar arteries and iliolumbar arteries. The patient was discharged in an improved clinical condition. Leriche's syndrome and pregnancy demonstrating complete aortic, common, and external iliac artery occlusion is very rare in the literature. Despite complete occlusion, viability of the fetus can be achieved with collateral vessels.

Regulatory effect of miR-195 in the placental dysfunction of preeclampsia.

Purpose: Preeclampsia (PE) is a pregnancy specific disease soon after 20 weeks of gestation where major symptoms are hypertension and proteinuria. The underlying pathology is believed to be abnormal placentation. Epigenetic and genetic factors have significant roles in abnormal placental development. MicroRNA's (miRNAs), being one of the most important epigenetic regulators, take part in abnormal placentation. Hsa-miR-195 is a molecule associated with abnormal placental growth mechanisms such as impaired cellular proliferation, inadequate trophoblastic invasion causing defective spiral artery remodeling, and apoptosis. We aimed to evaluate miRNA functions, namely miR-195 expression profile, in order to divulge PE pathogenesis.Methods: In this study, we extracted circulating miRNAs from maternal plasma and placenta from 20 PE patients and 20 normotensive pregnant women. miR-195 was quantified using quantitative real time reverse transcriptase PCR (qRT-PCR). The target genes of miR-195 were predicted by Diana Tools-mirPath, TargetScan, and miRDB databases.Results: We found that miR-195 levels were downregulated (3.83-fold decrease, p < .05) in preeclamptic placenta samples, however miR-195 were undetected in preeclamptic and normotensive plasma samples. The steep down-regulation of miR-195 points to its importance of PE pathogenesis.Conclusion: miR-195 is suggested to regulate PE via its target genes manipulating biological processes such as placental proliferation, apoptosis, and angiogenesis. We propose that detection of decreased miR-195 levels in preeclamptic placentas could be used to enlighten the pathophysiology of PE.

Regulation of HMGA2 and KRAS genes in epithelial ovarian cancer by miRNA hsa-let-7d-3p.

AIM OF THE STUDY: The purpose of this study was to identify specific circulating microRNAs (miRNAs) and investigate expression level of their target genes for evaluation of pathogenesis of epithelial ovarian cancer (EOC). MATERIALS AND METHODS: In this study, we have studied on EOC patients' serum and whole blood, healthy control (HC) serum, and whole blood samples. Sixteen serum samples were collected to compare miRNA expression analysis through microarray. According to microarray results, one of the dysregulated miRNA in serum, hsa-let-7d-3p, was validated by RT-qPCR for discriminate two groups. The hsa-let-7d-3p is one of the tumor suppressive let-7d family members. Let-7d is downregulated in numerous types of cancer, including ovarian cancer and directly targets various oncogenes. We analyzed the let-7d targets, which are High Mobility Group A2 (HMGA2) and (Kirsten Rat Sarcoma Viral Oncogene Homolog), as the oncogenes that are associated with EOC. The relation between target genes of hsa-let-7d-3p and EOC has been examined by Pathway Studio. Twenty serum and whole blood samples collected to analyze expression level of target genes were analyzed by real-time PCR. RESULTS: 31 significantly dysregulated miRNAs were identified by microarray in serum. Hsa-let-7d-3p has been selected for the validation, according to P-value and dysregulated level. RT-qPCR results showed that hsa-let-7d-3p could discriminate EOC patients from HC (P = 0.0484, AUC = 0.7). Furthermore, we identified hsa-let-7d-3p's target genes (HMGA2, KRAS) by bioinformatic analysis. The expression level of genes could discriminate patients with EOC from HC, with a power area under the ROC curves (AUC) of 62 and 64.2, respectively. CONCLUSION: HMGA2 and KRAS could be translationally downregulated by the hsa-let-7d-3p, and the loss of hsa-let-7d-3p expression led to the progression of EOC related to the tumorigenesis, invasion, and metastasis.

MicroRNA expression profiling in placenta and maternal plasma in early pregnancy loss.

Early pregnancy loss (EPL), also termed early miscarriage, is determined as the unintentional expulsion of an embryo or fetus prior to the 12th week of gestation. EPL frequency is ~15% in pregnancies. Fetal development and growth is associate with placental function and vessel development; therefore, the placental genome would represent a useful miscarriage model for (epi)genetic and genomic studies. An important factor of placental development and function is epigenetic regulation of gene expression. microRNAs (miRNAs) are the primary epigenetic regulators which have an important role in placental development and function. In the present study, maternal plasma and villous tissue were collected from 16 EPL cases in 6th­8th gestational weeks (GWs) and 8 abortions (control group) in 6th­8th GWs. Detection of the differences in miRNA expression was performed using microarrays and dysregulated miRNAs were validated by reverse transcription­quantitative polymerase chain reaction (RT­qPCR). miRNA microarray findings revealed that four miRNAs, including hsa­miRNA (miR)­125a­3p, hsa­miR­3663­3p, hsa­miR­423­5p and hsa­miR­575 were upregulated in tissue samples. In maternal plasma, two miRNAs (hsa­let­7c, hsa­miR­122) were upregulated and one miRNA (hsa­miR­135a) was downregulated. A total of 6 out of 7 dysregulated miRNAs were validated using RT­qPCR. The target genes of these dysregulated miRNAs were detected using the GeneSpring database. The aim of the present study was to detect dysregulated miRNAs in maternal plasma and villous cells and identify the target genes of dysregulated miRNAs and their associated pathways. The target gene analyses have revealed that the affected genes are primarily associated with cell migration, proliferation, implantation, adhesion, angiogenesis and differentiation and all are involved with EPL pathogenesis. Therefore, the present study may contribute to the understanding of the molecular mechanisms which lead to EPL.

Expression profiling of maternal plasma and placenta microRNAs in preeclamptic pregnancies by microarray technology.

Preeclampsia (PE) is one of the leading causes of maternal and fetal morbidity and mortality, occurring usually in the second half of pregnancy and affecting approximately 5-8% of pregnancies in the world. miRNAs play critical role in the regulation of placental development processes. We aimed to determine specific novel miRNAs for early diagnosis of preeclampsia which is one of the most dangerous pregnancy diseases. In this study 72 samples, maternal age 22 ≤ and ≤36, have been analyzed; maternal plasma and placental miRNAs were isolated from 18 severe preeclampsia (sPE) patients and 18 controls, respectively. Profiling of human miRNAs (1368 probe) was performed in samples with Agilent v16 microarrays for detection of the differences in miRNA expression between two groups. The results were validated by using TaqMan RT-qPCR method. The analysis indicated that 406 of these miRNAs in all placentas and 42 of these miRNAs in all maternal plasma were expressed. The relative expression analysis has shown that 12 miRNAs (p < 0.05 and >2-fold) in maternal plasma were differentially expressed in PE and control group. However, five miRNAs were validated by qRT-PCR. Once validated miRNAs have been searched in databases for their target genes and function, it has been shown that there are some preeclampsia related pathways as a target such as angiogenesis, cardiovascular, hypertension, placental abruption and preeclampsia disorders. Differentially expressed and validated plasma miRNAs might be used as notable biomarkers for non-invasive early diagnosis of preeclampsia and treatment of disease.

Serum leptin levels in hypertensive disorder of pregnancy.

OBJECTIVE: To determine serum leptin levels in hypertensive disorder of pregnancy. MATERIALS AND METHODS: In this prospective, cross-sectional, case control study, we measured serum leptin levels of 58 hypertensive pregnant women and 54 normal pregnant women. We also did blood and urine analysis for the evaluation of the severity of hypertensive disorder of pregnancy. The patients were followed until after delivery and information about labour was recorded. We analysed the difference and correlation between anthropometric measures, hormonal and biochemical parameters, and serum leptin levels in two groups. RESULTS: In the study group, serum leptin levels were determined to be higher than the control group. Neonatal birth weight was significantly lower in the hypertensive group. While the serum uric acid, urea, aspartate aminotransferase, fibronectin, and fasting blood glucose levels were found to be higher, serum total protein and albumin levels were significantly lower among the hypertensive pregnant women. Hypertensive pregnant women were more insulin resistant. Serum leptin levels were highly and positively correlated with serum fibronectin, and C peptide levels. A negative significant correlation was observed between maternal serum leptin levels and neonatal birth weight among the pregnant women having the hypertensive disorders. CONCLUSION: Serum leptin levels in hypertensive pregnant women appear to be higher. The determination of serum leptin levels may be as important as serum fibronectin and C peptide levels in the management of hypertensive disorder of pregnancy. C peptide and insulin may be due to hyperinsulinemia which leads to increased stimulation of leptin production by fatty tissue. Insulin resistance which appears in late pregnancy is more significant especially in pregnancies complicated by preeclampsia.

Significance of the O6-methylguanine-DNA methyltransferase and glutathione S-transferase activity in the sera of patients with malignant and benign ovarian tumors.

OBJECTIVE: To demonstrate O6-methylguanine-DNA methyltransferase (MGMT) and glutathione S-transferase (GST) activities by analyzing the sera separately obtained from patients with malignant ovarian tumors, benign ovarian tumors, and healthy individuals. STUDY DESIGN: Fourty-nine patients with ovarian cancer, nine patients with benign tumors, and 22 healthy women were included in this study. Blood samples were obtained from all the subjects in the malignant-tumor, benign-tumor, and control groups. Patients with malignant tumors underwent second and third phlebotomies one week following the surgery and after the chemotherapy regimen, respectively. MGMT, GST, and protein levels were measured for each serum sample. GST activity of the samples was measured by the method of Habig et al. using l-chloro-2-4 dinitrobenzene (CDNB) as substrate. MGMT activity was measured by the transfer of radio labelled methyl groups from a prepared MG-DNA substrate to the enzyme fraction of serum. Protein concentration was measured by biuret method. RESULTS: Our work demonstrated that untreated patients with malignant ovarian tumors revealed significantly greater MGMT and GST activities in their sera than did both healthy individuals and patients with benign ovarian tumors, while no significant difference was found between the healthy group and the patients with benign ovarian tumors with respect to their sera MGMT and GST activities. GST activity following chemotherapy was significantly lower than the postoperative values preceding chemotherapy. A relationship between sera MGMT and GST activities, tumor histology and pathology was not found in this study. CONCLUSION: Our work suggests the fact that detection of sera MGMT and GST activities is important in diagnostic and therapeutic approaches during the course of ovarian cancer.

Low-grade fibromyxoid sarcoma of the vagina: A tumor, not previously reported at this site.

This report presents the first case of low-grade fibromyxoid sarcoma (LGFMS) arising from the vaginal wall (a rare soft-tissue sarcoma of subfascial planes) and draws attention to differential diagnosis of masses arising from the vaginal wall. A patient presenting with abdominal pain to emergency department was diagnosed to have an ovarian mass filling the Douglas space. At laparoscopy, the origin of the mass was identified as the posterior vaginal wall. After vaginal excision of the gelatinous mass, pathologic diagnosis revealed a rare tumor, LGFMS. We discussed the differential diagnosis of vaginal LGFMS.

Effect of angiotensin I-converting enzyme and α-actinin-3 gene polymorphisms on sport performance.

Genetic polymorphism is considered to be associated with human physical performance. The angiotensin I-converting enzyme insertion/deletion (ACE I/D) and the α-actinin-3 gene (ACTN3) R577X polymorphisms have been widely investigated for such associations, and functional ACE I/D and ACTN3 R577X polymorphisms have been associated with sprinter performance. The aim of this study was to determine the effect of these polymorphisms on sport performance among 37 elite athletes and 37 healthy controls. The ACE II genotype was identified in 32.43% of the control group and 8.11% of elite athletes, the DD genotype in 37.84% of the control group and 51.35% of the elite athletes, and the ID genotype in 29.73% of the control group and 40.54% of the elite athletes. With regard to the ACTN3 gene, the XX genotype, which confers an advantage for endurance activities, was identified in 10.81% of the control group and 35.14% of the elite athletes. The XX genotype was observed more frequently than the RR genotype (advantageous for sprinting), which was identified in 2.70% of the control group and 10.81% of elite athletes. The RX genotype (observed in 86.48% of the control group and in 54.05% of the elite athletes) was the most common genotype of the individuals in the present study. The study showed that ACTN3 and ACE gene polymorphisms have an effect on muscle power; however, larger studies are required.

Current approaches on non-invasive prenatal diagnosis: Prenatal genomics, transcriptomics, personalized fetal diagnosis.

Recent developments in molecular genetics improved our knowledge on fetal genome and physiology. Novel scientific innovations in prenatal diagnosis have accelerated in the last decade changing our vision immensely. Data obtained from fetal genomic studies brought new insights to fetal medicine and by the advances in fetal DNA and RNA sequencing technology novel treatment strategies has evolved. Non-invasive prenatal diagnosis found ground in genetics and the results are widely studied in scientific arena. When Lo and colleges proved fetal genetic material can be extracted from maternal plasma and fetal DNA can be isolated from maternal serum, the gate to many exciting discoveries was open. Microarray technology and advances in sequencing helped fetal diagnosis as well as other areas of medicine. Today it is a very crucial prerequisite for physicians practicing prenatal diagnosis to have a profound knowledge in genetics. Prevailing practical use and application of fetal genomic tests in maternal and fetal medicine mandates obstetricians to update their knowledge in genetics. The purpose of this review is to assist physicians to understand and update their knowledge in fetal genetic testing from maternal blood, individualized prenatal counseling and advancements on the subject by sharing our experiences as Istanbul University Fetal Nucleic Acid Research Group.

Relationship between lymphocytes, IL2 and the hormones E2, LH, PRG and FSH in menopausal and postmenopausal women.

PROBLEM In this baseline study, our aim is to show the relationship of parameters and gonad hormones in menopausal and postmenopausal women. METHOD Blood samples were taken from menopausal and postmenopausal women (12-14 months and ≥10 years, respectfully, since their last menstruation). Adolescents aged 13.7 ± 0.7 were used as controls. Hormones were measured by ELISA and percentages of CD45, CD4, CD8, CD3, CD19, IL-2, CD25 and HLA-DR were measured by flow cytometry. RESULTS Both groups showed an increase in the percentage of CD3, CD4 and CD8. Levels of CD19 were significantly lower in the postmenopausal group. However, changes in immunologic parameters during menopause were less marked than the hormonal changes observed in these groups. Most of the correlations LH × CD3 (-ve), LH × IL2R (-ve) and E2 × CD19 (+ve) suggesting how menopausal women with particularly high LH or low E2 levels may be affected. Only CD3 and HLA-DR correlated with the hormonal changes in the postmenopausal group. IL-2 levels were high in the menopausal group and low in the postmenopausal group; however, no correlation was observed. DISCUSSION Menopause is characterized by increased levels of IL-2, which has critical immune-modulatory effects. These changes may be related to the overall hormonal change process observed during menopause.

Comparison of real-time polymerase chain reaction assay methods for detection of RHD gene in amniotic fluid.

Hemolytic disease of the newborn is the clinical condition in which Rh blood group antigens in couples are incompatible with each other and mother is negative for the antigen, whereas father is positive. Although RHD antigen encoded by RHD gene that is localized on chromosome 1 determines person's Rh genotyping, this incompatibility can lead to delivery as anemia, jaundiced, or dead in mother's uterus. In recent years, improvements have occurred in the prenatal diagnosis of Rh incompatibility. Quantitative real-time polymerase chain reaction (Real-time PCR) has been improved and determining rapidly, reliably, and sensitively has been possible. In this study, the determination of RHD genotyping was investigated using fetal DNA obtained from amniotic fluid and SYBR Green I and TaqMan probe methods were compared, and reliability in prenatal diagnosis of these methods was determined. We studied 35 pregnant women in the second trimester of pregnancy. "SYBR Green I" and "TaqMan" probes results for RHD gene of genomic DNA extracted from total 35 different amniotic fluid samples acquired from 10 RHD (-) and 25 pregnant women randomly were analyzed. DNA extracted from amniotic fluid was analyzed for RHD gene with real-time PCR and the results were then compared with the RHD fetal genotype determined on RHD phenotype of the red blood cells of the infants at birth. The results of RHD TaqMan probes PCR analysis of amniotic fluid DNA were completely concordant with the fetal blood group analysis after birth. Real-time PCR using the TaqMan probes has proven to be more sensitive, accurate, and specific for RHD gene than SYBR Green I method.

Detection of fetal RhD gene from maternal blood.

OBJECTIVE: Hemolytic disease of the newborn (HDN) is a clinic phenomenon which occurs during pregnancy due to the Rhesus (Rh) D alloimmunization between a Rh (-) pregnant woman, who has become sensitive to RhD antigens, and her Rh (+) fetus. As a result of the attack of maternal RhD antibodies on fetal RhD antigens, fetal anemia, HDN and fetal death may occur. % 40 of Rh (-) pregnant women carry Rh (-) fetus. However, all Rh (-) pregnant women are offered anti-D Immunoglobulin (Anti-D Ig) at 28 weeks' gestation in case of fetomaternal haemorrhage, so the pregnant women carrying Rh (-) fetus are exposed to blood products unnecessarily. Although the RhD of fetus can be detected, methods used for prenatal diagnosis recently are invasive tests and they can result in abortion in a certain percentage. The discovery of circulating cell-free fetal nucleic acids in maternal plasma has opened up new possibilities for non invasive prenatal diagnosis. The aim of this study was to detect prenatal RhD by analysing the presence of the RhD gene of fetal DNA in maternal blood. MATERIAL AND METHODS: Total free DNA was isolated from the blood of 19 Rh (-) pregnant women, who had RhD alloimmunization with their husbands, in the 11-14 th week of their pregnancy. The existence of a gene in isolated DNA was investigated with TaqMan prob and "Real-time PCR" method by using primers belonging to exon 7 of RhD gene. RESULTS: Using a quantitative real-time PCR assay, the presence of RhD gene sequences was evaluated in the serum of patients at the onset of pregnancy. We have analyzed 19 Rh (-) pregnant women. Twelve of them were Rh (-) and the rest of them were 7 Rh (+). After birth the baby's blood groups were concordant with our results. CONCLUSION: The results obtained by RhD primer were analysed. The possibility of detection of fetal RhD gene in maternal blood contributed to noninvasive prenatal diagnosis.