Prevalence of 1691G>A FV mutation in Poland compared with that in other Central, Eastern and South-Eastern European countries

  • Grażyna Adler Department of Medical Biology, Pomeranian Medical University
  • Jeremy S C Clark Department of Clinical and Molecular Biochemistry, Pomeranian Medical University
  • Beata Łoniewska Department of Neonatal Diseases, Pomeranian Medical University
  • Ewa Czerska Central Laboratory, Regional Hospital
  • Nermin N Salkic Department of Gastroenterology and Hepatology, University Clinical Center Tuzla
  • Andrzej Ciechanowicz Department of Clinical and Molecular Biochemistry, Pomeranian Medical University
Keywords: Factor V, FV Leiden, centroids

Abstract

The 1691G>A FV variant has been described as a common genetic risk factor in venous thromboembolism. The purpose of this study was to provide a further frequency value for 1691G>A FV in Poland and to collate summary data from Central (Poland, Czech, Slovakia), Eastern (Russia, Belarus, Ukraine) and South-Eastern (Slovenia, Croatia, Bosnia and Herzegovina, Serbia, Montenegro, Macedonia, Bulgaria) European countries. For this purpose in 2007 the 1691G>A FV variant was analyzed by polymerase chain reaction-restriction fragment length polymorphism from DNA collected in 2005-2006. We studied 650 subjects: 400 newborns and 250 older individuals (mean age 46.1 y) from Poland and compared results with reports from other countries, as well as with the frequency trend of 845G>A HFE across South-Eastern European countries using centroid cities. From our 1691G>A FV study we identified 626 GG homozygotes, 23 GA heterozygotes, and i AA homozygote (n = 650), giving an A allele frequency of 1.9%, and a summed frequency value for Poland of 2.0% (n = 1588); the frequency in Central European countries was 3.9% (n = 4559), mostly due to the high value in the Czech Republic: 5.1% (n = 2819); the South-Eastern European countries had 2.5% (n = 2410). Among the Eastern European countries the 1691G>A FV allele frequency was 1.9% (n=791), between the South-Eastern and Eastern European countries there was no significant difference (p=0.17). We confirm that the 1691G>A FV allele frequency in Poland, as well as other countries compared, is significantly lower than that in Czech.

Downloads

Download data is not yet available.
Prevalence of 1691G>A FV mutation in Poland compared with that in other Central, Eastern and South-Eastern European countries
Published
2012-05-20
How to Cite
1.
Adler G, S C Clark J, Łoniewska B, Czerska E, N Salkic N, Ciechanowicz A. Prevalence of 1691G>A FV mutation in Poland compared with that in other Central, Eastern and South-Eastern European countries. Bosn J of Basic Med Sci [Internet]. 2012May20 [cited 2021Oct.18];12(2):50-4. Available from: https://www.bjbms.org/ojs/index.php/bjbms/article/view/2500
Section
Molecular Biology

INTRODUCTION

Synthesized in the liver, blood coagulation Factor V (FV) is a multidomain glycoprotein encoded by a gene consisting of 25 exons, located on chromosome 1q23. The 1691G>A FV transition in exon 10 of factor 5 causes an arginine to glutamine substitution (R506Q) known as Factor V Leiden. This genetic disorder is characterized by poor anticoagulant response to activated protein C and is the most common risk factor for thromboembolic disease [1]. Major clinical observations are that the presence of 1691G>A FV increases risk of deep vein thrombosis [2-4] and is also associated with a increased relative risk for pregnancy loss and possibly other obstetric complications [5,6].

The frequency of the 1691G>A FV allele varies worldwide and differences are observed between geographic locations and ethnic populations: The 1691G>A FV allele is very rare or non-existent in Asia (0.6%) and some regions of Africa (0.0%) [7-9]. On the other hand, Settin et al. [10] has described the prevalence of the mutant allele at the level of 10.2% in Egypt. In Poland, the prevalence of the variant 1691G>A FV has been previously given by several researchers [11-15]. One objective of this research was to give a larger sample size with a 650 subjects from the West Pomeranian province of Poland. This value is then compared to previous groups of Poles and values from other countries. Note that the population now inhabiting the region of West Pomerania resulted from extensive mixing of Polish peoples from all regions of Poland after the Second World War and therefore can provide a representative sample for the whole of Poland [16, 17]. A second objective was to present summary data: to our knowledge summary data for 1691G>A FV from Central (Poland, Czech, Slovakia), Eastern (Russia, Belarus, Ukraine) and South-Eastern (Slovenia, Croatia, Bosnia and Herzegovina, Serbia, Montenegro, Macedonia, Bulgaria) European countries has not been presented before. The third aim of this study was to provide summed frequency values for 1691G>A FV in these countries, gathered from studies using similar methods ie. by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

MATERIAL AND METHODS

Samples

The experimental study was performed (in 2007) on a group of 650 Polish individuals, divided into two subgroups: 400 newborns (187 female and 213 male) - and 250 older subjects (mean age 46.1 y, range: 2-87 y, 169 female and 81 male) - all inhabitants of the West Pomeranian province of Poland. The older subjects, of Polish origin, were consecutive healthy visitors to the analytical laboratory VITA at Darlowo, Poland, and no exclusion criteria were used. The sub-group of newborns has been described previously [14]. All neonates were of Polish origin, with Polish grandparents, and informed consent was obtained from all parents. The Ethical Committee of the Pomeranian Medical University approved the protocol of the study (BN-001/57/05).

Procedure

For identification of the NM_000130.4:c.1691G>A F5 alteration (here designated as 1691G>A FV) we used PCR-RFLP. Genomic DNA was extracted from 100 μL of umbilical blood (for newborns) or full blood (for older subjects), using the QIAamp DNA Blood Mini Kit (QIAGEN, Hilden, Germany). For a 10 μl-PCR, ~20 ng of genomic DNA was used. The PCR mixture contained 10x buffer (pH 8.3, 1.5 mM MgCl2), 0.2 mM each of the deoxynucleoide triphosphates, 0.5 U Polymerase Taq (MBI Fermentas, Lithuania) and 4 pmol each of the forward and reverse primers as designed by Gandrille et al. [18]. Primers were synthesized by TIB MOLBIOL, Poznan, Poland. PCRs were performed in a Mastercycler Gradient device (Eppendorf, Hamburg, Germany) with the following temperature profile: initial denaturation at 94°C for 5 min; 35 cycles of 20 s at 94°C, 40 s at 56°C and 40 s at 72°C; and final extension step at 72°C for 7 min. Amplification was followed by digestion of a 241 bp product with Hind III restriction enzyme (5 ‘-A↓AGCTT-3’) (MBI Fermentas, Lithuania) for 16 hours at 37°C. The PCR digestion products were separated in 3 % agarose gels, stained with ethidinum bromide and recorded with a DS-34 Polaroid Instant Camera (Polaroid, Germany) using UV light (Transilluminator 4000, Stratagene, La Jolla, CA, USA). Hind III digestion yields fragments: 241 bp. (homozygote GG); 241, 209 and 32 bp. (heterozygote GA); 209 and 32 bp. (homozygote AA). Genotypes of GA and AA subjects were also confirmed by DNA sequencing (3100-Avant Genetic Analyzer, Applied Biosystems Hitachi, Foster City, CA, USA). For summary trend data of 1691G>A FV in South-Eastern European countries including Turkey the values are plotted against latitude of centroid cities (from BRRG - Buero fuer Raumforschung, Raumplanung und Geoinformation, Oldenburg, Germany: http://www.brrg.de/database. php?language= en&cId=0&dId=47) with latitude derived from Google maps (Google Inc, USA; http://www.mapcrow.info): Centroid cities “represent the political, administrative and cultural centre of the region”. Graphical materials were developed using Designworks software (GSP Ltd, London, UK) and Microsoft Office (Microsoft, Redmond, WA, USA).

Statistical analysis

Fifty-five statistical comparisons (using z-tests) were made between all pairs of summary prevalence values for all countries studied. With Bonferroni correction a critical p-value of (0.05/55 = 0.0009) was used to define statistical significance. Z-tests and correlation coefficients (tested by linear regression) were calculated using Statistica (data analysis software system, version 8.0, StatSoft, Inc. 2007, www.statsoft.com).

RESULTS

The frequency of the 1691G>A FV allele in the study group (n = 650) proved to be 1.9%. We identified 626 GG homozygotes, 23 GA heterozygotes, and 1 AA homozygote, conforming to the expected Hardy-Weinberg equilibrium. This gives a summed frequency value for Poland of 2.0% (n = 1588). The average frequency of the 1691G>A FV allele in Central, Eastern and South-Eastern European countries was 3.2 % and varied from 0.6% (Belarus) to 5.1% (Czech Republic). The frequency of 1691G>A FV from our results (Table 1) and summary data from other countries (Table 2) are shown, and a map showing the summed frequencies for 1691G>A FV is shown in Figure 1. In Central European countries the frequency of 1691G>A FV was 3.9% (n = 4559) and varied from 5.1% in the Czech Republic through 2.1% in Poland (including our study) to 1.3% in Slovakia [11-13, 15, 19, 20]. The Czech Republic value was found to be significantly different from that in Poland, Russia, Ukraine, Slovenia, Croatia and Serbia/Montenegro, using two-proportion z-tests for comparisons between all countries studied except Turkey (with Bonferroni correction,p< 0.001, Table 3). In Eastern European countries the frequency of this mutant allele was 1.9% (n = 791) and varied from 2.4% in Russia through 0.9% in Ukraine to 0.6% in Belarus [43, 45, 46]. The prevalence of the 1691G>A FV allele in South-Eastern European countries was 2.5% (n=2410) varying from 3.6% in Bulgaria to 1.6% in Croatia. Unfortunately no data were found for Bosnia and Herzegovina, despite an extensive search. The 1691G>A FV variant follows a roughly increasing trend from West to East (Figure 2).

TABLE 1: Allele frequencies for 1691G>A FV in present study.
TABLE 2: Allele frequencies for 1691G>A FV in Central, Eastern and Southeastern European countries.
FIGURE 1: Allele frequencies (bold), number of subjects (not bold) for 1691G>A FV in Central, Eastern and South-Eastern countries (summed frequencies from references in Table 2).
TABLE 3: p-values from two-proportion z-tests between 1691A FV frequency values of Central, Eastern and South-Eastern European countries. Significant differences, after Bonferroni correction (critical p = 0.05/55 = 0.0009), are shown in bold.
FIGURE 2: Trend in frequency of 1691G>A FV in South-Eastern Europe. Centroid cities: 1, Ljubljana (Slovenia); 2, Zagreb (Croatia); 3, Belgrade (Serbia and Montenegro); 4, Skopje (Macedonia); 5, Sofia (Bulgaria); 6, Istanbul (Turkey)

DISCUSSION

In the countries examined, which are predominantly inhabited by Slavic peoples, the allele frequencies for 1691G>A FV provide a mosaic (Figure 1). In our study group the frequency of the 1691G>A FV allele was consistent with the previous summed frequency value for Poland (2.0%) [11-13,15] and is similar to that in France (2.2%) [47], Switzerland [48] and the Netherlands [49-50] (each 2.2%) as well as to that in Serbia (2.2%) [31-35] and Russia (2.4%) [43, 44]. This value is, however, significantly different from that in the adjacent Czech Republic. To our knowledge we give the first summary study of the frequency distribution of the 1691G>A FV allele in Central, Eastern and South-Eastern European countries. A total of 7760 control individuals originating from 11 countries in provide the value for the frequency (3.2%) of this mutated allele in these countries. However, if the Czech value is removed the summed frequency for the 10 remaining countries is 2.2%. These percentages indicate that future genetic counseling will be of some benefit in this region. The reasons for this mosaic in 1691G>A FV frequencies is not known. However, population movements have contributed to ethnic groups, cultures and consequently inheritance mixing, both throughout thousands of years of prehistory as well as in recent documented history. In the Southern Slavs there is a rough upward trend for 1691G>A FV in a southeasterly direction from Croatia (1.6%) and Serbia (2.2%) to Macedonia (3.5%) and Bulgaria (3.6%) (Figure 2). In neighboring Turkey this trend continues in a southeasterly direction as the frequency is even higher, at 5.0% [51-57]. This trend opposes that for 845G>A HFE [58]. It would be interesting to fill the gaps in the data for 1691G>A FV in Bosnia and Herzegovina, and it would be interesting to examine the prevalence of 1691G>A FV in the Western Slavic group in Germany (the Sorbs).

CONCLUSIONS

The frequency of the 1691G>A FV allele in Poland, summed from our study and previous studies, 2.0%, was similar to that in most countries studied, and similar to the summed frequency value for all Central, Eastern and South-Eastern countries (with the value for the Czech Republic removed), ie. 2.2%. The values for Poland, Russia, Ukraine, Slovenia, Croatia and Serbia/Montenegro were significantly different from that in the adjacent Czech Republic. In the South-Eastern European countries there is a rough upward trend for 1691G>A FV in a southeasterly direction, which opposes that for 845G>A HFE (Figure 2 [58]).

Acknowledgements

ACKNOWLEDGEMENT

Funding: This study was funded entirely by the Pomeranian Medical University, Szczecin, Poland. We would like to thank César de Diego Díez, at GISCO, Luxembourg, for providing NUTS population centroid data.

DECLARATION OF INTEREST

The authors state there is no conflict of interest.

REFERENCES

  1. (). Factor V Leiden thrombophilia. Genet Med.
  2. , , , , , (). Venous thrombosis due to poor anticoagulant response to activated protein C: Leiden Thrombophilia Study. Lancet.
  3. , , , , , (). Factor V Leiden and risks of recurrent idiopathic venous thromboembolism. Circulation.
  4. , (). Activated protein C resistance and factor V Leiden: a review. Arch Pathol Lab Med.
  5. , , , , , (). Factor V Leiden variant as a risk factor for recurrent pregnancy loss. Ann Intern Med.
  6. , , , , , (). Factor V leiden and prothrombin G20210A variants, but not methylentetrahydrofolate reductase C677T, are associated with recurrent miscarriages. Hum Reprod.
  7. , , , , (). Polymorphisms in various coagulation genes in black South African women with placental abruption. BJOG.
  8. , , , , (). Impact of thrombogenic mutations on clinical phenotypes of von Willebrand disease. Clin Appl Thromb Hemost.
  9. , , , , , (). Prevalence of angiotensin-converting enzyme, methylenetetrahydrofolate reductase, Factor V Leiden, prothrombin and apolipoprotein E gene polymorphisms in Morocco. Ann Hum Biol.
  10. , , , , , (). Frequency of factor V Leiden variant in Egyptian cases with myocardial infarction. Hematology.
  11. , , , , , (). Prevalence of factor V Leiden variant in various populations. Genet Epidemiol.
  12. , , , , , (). Factor V Leiden, prothrombin gene G20210A variant, and methylenetetrahydrofolate reductase C677T genotype in young adults with ischemic stroke. Clin Appl Thromb Hemost.
  13. , , , , , (). Genetic polymorphisms associated with acute pulmonary embolism and deep venous thrombosis. Eur Respir J.
  14. , , , , , (). An age-related decrease in factor V Leiden frequency among Polish subjects. J Appl Genet.
  15. , , , (). The significance of genetic polymorphisms of factor V Leiden and prothrombin in the preeclamptic Polish women. J Thromb Thrombolysis.
  16. , , , , , (). Homogeneity and distinctiveness of Polish paternal lineages revealed by Y chromosome haplogroup analysis. Human Genetics.
  17. , , , , , (). Significant genetic differentiation between Poland and Germany follows present-day political borders, as revealed by Y-chromosome analysis. Human Genetics.
  18. , , (). A rapid screening method for the factor V Arg506-->Gln variant. Blood Coagul Fibrynolysis.
  19. , , , , (). Factor V Leiden in pregnancies complicated by placental abruption. BJOG.
  20. , , , , , (). The factor V Leiden variant in users of hormonal contraceptives. Ceska Gynekol.
  21. , , , , , (). Factor V Leiden and the Slovak population [in Czech]. Vnitr Lek.
  22. , , , , , (). Factor V Leiden, prothrombin 20210G >A, methylenetetrahydrofolate reductase 677C >T and plasminogen activator inhibitor 4G/5G polymorphism in women with pregnancy-related venous thromboembolism. Eur J Obstet Gynecol Reprod Biol.
  23. , , , , , (). Prothrombotic gene polymorphisms and atherothrombotic cerebral infarction. Acta Neurol Scand.
  24. , , , (). Joint effect of Gc.1691G>A factor V point variant and factor 5II Arg/Gln(353) gene polymorphism on the risk of premature coronary artery disease. Ann Genet.
  25. , , , (). Major and potential prothrombotic genotypes in patients with venous thrombosis and in healthy subjects from Slovenia. Pathophysiol Haemost Thromb.
  26. , , , , (). Prevalence and association of the factor V Leiden and prothrombin G20210A in healthy subjects and patients with venous thromboembolism. Croat Med J.
  27. , , , , , (). ABO blood groups and genetic risk factors for thrombosis in Croatian population. Croat Med J.
  28. , , , , , (). Prevalence of factor V Leiden and G6PD 1311 silent variants in Dalmatian population. Arch Med Res.
  29. , , , , , (). Frequency of factor II G20210A, factor V Leiden, MTHFR C677T and PAI-1 5G/4G polymorphism in patients with venous thromboembolism: Croatian case-control study. Bioch Med.
  30. , , , , (). Lower contribution of factor V Leiden or G202104 variants to ischemic stroke in patients with clinical risk factors: pair-matched case-control study. Clin Appl Thromb Hemost.
  31. , , , , , (). Thrombophilia in women with pregnancy-associated complications: fetal loss and pregnancy-related venous thromboembolism. Gynecol Obstet Invest.
  32. , , , (). Prevalence of factor V Leiden variant in Yugoslav thrombophilic patients and its relationship to the laboratory diagnosis of APC resistance. Thromb Haemost.
  33. , , , , , (). Prevalence of factor V leiden, factor V cambridge, factor II G20210A and methylenetetrahydrofolate reductase C677T variants in healthy and thrombophilic Serbian populations. Acta Haematol.
  34. , , , , (). FV Leiden variant and deep venous thrombosis in Vojvodina: a case-control study. J Med Biochem.
  35. , , , , , (). The FV Leiden, FII G20210A and MTHFR C677T variants in healthy and thrombophilic Yugoslav population. J Thromb Haemost. Abstr: P0338
  36. , , (). Factor V Leiden is associated with higher risk of deep venous thrombosis of large blood vessels. Croat Med J.
  37. , , , , (). Prevalence of factor V Leiden and prothrombin 20210 A variant in Bulgarian patients with pulmonary thromboembolism and deep venous thrombosis. Blood Coagul Fibrinolysis.
  38. , , , , (). Genetic thrombophilic defects (Factor V Leiden, prothrombin G20210A, MTHFR C677T) in women with recurrent fetal loss [in Bulgarian]. Akush Ginekol (Sofia).
  39. , , , , (). Carriers of thrombophilic factor among women with preeclampsia (preliminary report) [in Bulgarian]. Akush Ginekol (Sofia).
  40. , , , , , (). Impact of thrombophilic genetic factors on pulmonary embolism: early onset and recurrent incidences. Lung.
  41. , , , , , (). Inherited thrombophilic factors in women with unexplained intrauterine fetal deaths [in Bulgarian]. Akush Ginekol (Sofia).
  42. , , , , , (). Association of inherited thrombophilia with embryonic and postembryonic recurrent pregnancy loss. Blood Coagul Fibrinolysis.
  43. , , , , , (). Age as a risk factor for myocardial infarction in Leiden variant carriers. Mol Genet Metab.
  44. , , , , , (). C677T variant in methylentetrahy-drofolatereductase gene in patients with venous thromboses from the central region of Russia correlates with a high risk of pulmonary artery thromboembolism [in Russian]. Ter Arkh.
  45. , , (). Allelic polymorphism of FII, FV and MTHFR genes in population of Ukraine. Tsitol Genet.
  46. , , (). Thrombotic complications during cancer treatment in children. Exp Oncol.
  47. , , , , , (). Site of venous thromboembolism and prothrombotic mutations according to body mass index. Results from the EDITH study. Br J Haematol.
  48. , , , , , (). Coagulation factors II, V, VII, and X, prothrombin gene 20210G-->A transition, and factor V Leiden in coronary artery disease: high factor V clotting activity is an independent risk factor for myocardial infarction. Arterioscler Thromb Vasc Biol.
  49. , , , , , (). Prothrombotic conditions, oral contraceptives, and the risk of ischemic stroke. J Thromb Haemost.
  50. , , , , (). Gender-specific association of the factor V Leiden variant with fertility and fecundity in a historic cohort. The Leiden 85-Plus Study. Hum Reprod.
  51. , (). Frequency of factor V Leiden in Turkey. Int J Hematol.
  52. , , , , , (). Frequency of Factor V (1691 G>A) variant in Turkish population. Thromb Haemost.
  53. , (). The prevalence of factor V Leiden (1691 G- ->A) variant in Turkey. Turk J Pediatr.
  54. , , , , (). Factor V Leiden and Prothrombin gene 20210A variant in neonatal thromboembolism and in healthy newborns and adults: a study in a single center. Pediatr Hematol Oncol.
  55. , , , , (). Prevalence of prothrombotic abnormalities in patients with acute mesenteric ischemia. World J Surg.
  56. , , , , , (). The prevalence of factor V Leiden gene variant analysis of donor and recipient at the organ transplantation Center of Ege University. Ege Tip Dergisi.
  57. , , , , , (). Frequency of factor V Leiden (Gc.1691G>A), prothrombin (G20210A) and methylenetetrahydrofolate reductase (C677T) genes variants in woman with adverse pregnancy outcome. J Turkish-German Gynecol Assoc.
  58. , , , (). Prevalence of 845G>A HFE mutation in Slavic populations: an east-west linear gradient in South Slavs. Croat Med J.