Int J Lab Hem. 2017;1–6. wileyonlinelibrary.com/journal/ijlh | 1 © 2017 John Wiley & Sons Ltd Received: 2 March 2017 | Accepted: 24 April 2017 DOI: 10.1111/ijlh.12703 ORIGINAL ARTICLE Thrombophilic gene polymorphisms and recurrent pregnancy loss in Greek women M. Chatzidimitriou 1 | D. Chatzidimitriou 2 | M. Mavridou 1 | C. Anetakis 1 | F. Chatzopoulou 2 | T. Lialiaris 3 | S. Mitka 1 1 Medical Laboratories, Alexandre Technological Institute of Thessaloniki, Thessaloniki, Greece 2 Medical School, Aristotle University of Thessaloniki, Thessaloniki, Greece 3 Medical School, Demokritus University of Thrace, Thessaloniki, Greece Correspondence Maria Chatzidimitriou, Medical Laboratories, Alexandre Technological Institute of Thessaloniki, Thessaloniki, Greece Emails: chatzidi@med.auth.gr, mchatzid952@ gmail.com Abstract Introduction: Recurrent pregnancy loss (RPL) is a multifactorial disorder. The aim of this study was the detection of various genetic polymorphisms and their correlation to RPL, in Greek women. Methods: The impact of 12 thrombophilic polymorphisms was evaluated, among 48 Greek women with a history of RPL, vs 27 healthy parous women. Multiplex PCR and in situ hybridization on nitrocellulose films were performed, to investigate 12 genetic polymorphisms previously reported as risk factors for RPL. Results: Heterozygous FV Leiden, homozygous PAI-1 4G/4G, heterozygous MTHFR C677T, homozygous MTHFR A1298C, as much as the combined thrombophilic geno- types MTHFR 677T + ACE Ι/D, MTHFR 677T/1298C + ACE D/D, ACE I/D + b- fibrinogen -455 G/A, FV HR2 + b-fibrinogen -455 G/A showed a correlation as risk factors for RPL, whereas the rest of the investigated polymorphisms and their combi- nations did not render statistically significant differences between the two groups in study. Conclusion: The results of this study, as well as those of similar studies, concerning the detection of genetic, environmental, and physiological factors underlying RPL, will prove of critical significance in the investigation and treatment of thrombophilic pre- disposition, in cases of RPL. KEYWORDS gene polymorphisms, genetic risk factors, in situ hybridization, recurrent pregnancy loss, thrombophilia 1 | INTRODUCTION Recurrent pregnancy loss (RPL) is the occurrence of two or more con- secutive fetal loss prior to 20 weeks gestation or at the time when the fetus weighs less than 500 g. 1 Spontaneous abortion is the most com- mon complication of pregnancy, affecting approximately 10%-15% of clinically recognized pregnancies. 2 As much as RPL is one of the most common reproductive complications, it is also one of the least under- stood pathophysiological processes.. 3 As there is no known cause for 50% to 75% of this condition, the involvement of genetic risk factors is postulated to contribute in the etiology of RPL. 4 Pregnancy is a hypercoagulable state characterized by increased levels of prothrombotic factors and decreased antithrombotic fac- tors. Hemostatic failures may cause an obstruction of the placental circulation; hence, unexpected changes in coagulation factors during pregnancy may be the reason for RPL. 5 As of late, the association of RPL with maternal thrombophilic or hypofibrinolytic gene variants has assembled growing evidence. 6 Such thrombophilias augment the prothrombotic state of pregnancy, lead to inadequate fetomaternal circulation, and affect the process of placentation in the developing embryo. 2 In literature, twelve thrombophilic gene variants of ten ge- netic loci are largely associated with an increased risk of pregnancy