Iranian Journal of Neonatology

Iranian Journal of Neonatology

Correlation of Maternal Age, Weight, Pregnancy - associated Plasma Protein A, Free Beta-human Chorionic Gonadotropin, Fetal Crown-rump Length, and Fetal Gender with Fetal DNA Fraction in Non-invasive Prenatal Testing: An Experiment on Iranian Pregnant Women

Document Type : Original Article

Authors
Milad Dolatkhah 1
Marjan Rahnamaye Farzami 2
Ramezan-Ali Khavari-Nejad 1
Shokoofeh Noori 3
1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 Reference Health Laboratory, Ministry of Health and Medical Education, Tehran, Iran
3 Department of Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
10.22038/ijn.2020.45840.1765
Abstract
Background: Non-invasive prenatal testing (NIPT) as a novel screening method has been widely proposed to screen for common fetal chromosomal aneuploidies. The aim of the present study was to examine the possible effects of maternal age, maternal weight, fetal crown-rump length (CRL), serum pregnancy-associated plasma protein A (PAPP-A), free beta-human chorionic gonadotropin (free β-hCG), and fetal gender on cell-free fetal DNA (cffDNA) percentage fluctuations.
Methods: In the present cross-sectional study, 308 singleton pregnant women aged 20-47 years at 11+0 to 13+6 weeks of pregnancy referring to the DeNA Laboratory in Tehran, Iran, for NIPT test during a one-month period between July 2018 and August 2018 were selected randomly. The cffDNA was extracted from maternal plasma. Whole exome sequencing by a ion semiconductor sequencer using cffDNA was applied for all participants. The PAPP-A and free β-hCGas biochemical biomarkers were assessed using a closed chemilumine scence immunoassay analyzer. The Shapiro-Wilk test, Pearson analysis, beta regression analysis, and Mann-Whitney U test were performed to analyze the data.
Results: In the screening population, the cffDNA percentage showed no significant correlation with CRL and maternal age (P=0.096 and P=0.881, respectively). However, the cffDNA percentage correlated well with maternal weight, PAPP-A, and free β-hCG (P=0, P=0.009, and P=0.001, respectively). Beta regression between cffDNA percentage and maternal weight, free β-hCG, and PAPP-A was significant (P<0.001). The mean cffDNA percentage between male and female fetal groups showed a significant difference (P<0.001).
Conclusion: This study demonstrated that the cffDNA percentage in the first trimester of pregnancy had a negative correlation with maternal weight and a positive correlation with PAPP-A and free β-hCG values. Furthermore, the cffDNA percentage in male fetuses was higher than that in female fetuses.
 
 
Keywords
Cell-free nucleic acids
Chorionic gonadotropin
Pregnancy-associated plasma protein-A
Prenatal diagnosis
  1. VIncent MC, Daudin M, De MP, Massat G, Mieusset R, Pontonnier F, et al. Cytogenetic investigations of infertile men with low sperm counts: a 25‐year experience. J Androl. 2002; 23(1):18-22.
  2. Corrado F, Cannata M, Galia TL, Magliarditi M, Imbruglia L, D'anna R, et al. Pregnancy outcome following mid-trimester amniocentesis. J Obstet Gynaecol. 2012; 32(2):117-9.
  3. Lee FK, Chen LC, Cheong ML, Chou CY, Tsai MS. First trimester combined test for Down syndrome screening in unselected pregnancies-a report of a 13-year experience. Taiwan J Obstet Gynecol. 2013; 52(4):523-6.
  4. Li WH, Wang PH, Chuang CM, Chang YW, Yang MJ, Chen CY, et al. Noninvasive prenatal testing for fetal trisomy in a mixed risk factors pregnancy population. Taiwan J Obstet Gynecol. 2015; 54(2):122-5.
  5. Xue Y, Zhao G, Li H, Zhang Q, Lu J, Yu B, et al. Non-invasive prenatal testing to detect chromosome aneuploidies in 57,204 pregnancies. Mol Cytogenet. 2019; 12(1):29.
  6. Liao GJ, Gronowski AM, Zhao Z. Non-invasive prenatal testing using cell-free fetal DNA in maternal circulation. Clin Chim Acta. 2014; 428:44-50.
  7. Volik S, Alcaide M, Morin RD, Collins C. Cell-free DNA (cfDNA): clinical significance and utility in cancer shaped by emerging technologies. Mol Cancer Res. 2016; 14(10):898-908.
  8. Swanson A, Sehnert AJ, Bhatt S. Non-invasive prenatal testing: technologies, clinical assays and implementation strategies for women’s healthcare practitioners. Curr Genet Med Rep. 2013; 1(2):
    113-21.
  9. Qiao L, Zhang Q, Liang Y, Gao A, Ding Y, Zhao N, et al. Sequencing of short cfDNA fragments in NIPT improves fetal fraction with higher maternal BMI and early gestational age. Am J Transl Res. 2019; 11(7):4450-9.
  10. Barrett AN, Xiong L, Tan TZ, Advani HV, Hua R, Laureano-Asibal C, et al. Measurement of fetal fraction in cell-free DNA from maternal plasma using a panel of insertion/deletion polymorphisms. PLoS One. 2017; 12(10):e0186771.
  11. Canick JA, Kloza EM, Lambert‐Messerlian GM, Haddow JE, Ehrich M, van den Boom D, et al. DNA sequencing of maternal plasma to identify Down syndrome and other trisomies in multiple gestations. Prenat Diagn. 2012; 32(8):730-4.
  12. Dan S, Wang W, Ren J, Li Y, Hu H, Xu Z, et al. Clinical application of massively parallel sequencing‐based prenatal noninvasive fetal trisomy test for trisomies 21 and 18 in 11,105 pregnancies with mixed risk factors. Prenat Diagn. 2012; 32(13):1225-32.
  13. Liu S, Chen L, Zhang X, Li J, Lin H, Liu L, et al. Primer-introduced restriction analysis polymerase chain reaction method for non-invasive prenatal testing of β-thalassemia. Hemoglobin. 2015; 39(1):18-23.
  14. Zhang H, Gao Y, Jiang F, Fu M, Yuan Y, Guo Y, et al. Non‐invasive prenatal testing for trisomies 21,18 and 13: clinical experience from 146,958 pregnancies. Ultrasound Obstet Gynecol. 2015; 45(5):530-8.
  15. Chaudhuri K, Su LL, Wong PC, Chan YH, Choolani MA, Chia D, et al. Determination of gestational age in twin pregnancy: which fetal crown–rump length should be used? J Obstet Gynaecol Res. 2013; 39(4):761-5.
  16. Lun FM, Chiu RW, Chan KA, Leung TY, Lau TK, Lo YM. Microfluidics digital PCR reveals a higher than expected fraction of fetal DNA in maternal plasma. Clin Chem. 2008; 54(10):1664-72.
  17. Wang E, Batey A, Struble C, Musci T, Song K, Oliphant A. Gestational age and maternal weight effects on fetal cell‐free DNA in maternal plasma. Prenat Diagn. 2013; 33(7):662-6.
  18. Palomaki GE, Kloza EM, Lambert-Messerlian GM, Haddow JE, Neveux LM, Ehrich M, et al. DNA sequencing of maternal plasma to detect Down syndrome: an international clinical validation study. Genet Med. 2011; 13(11):913-20.
  19. Farina A, LeShane ES, Lambert-Messerlian GM, Canick JA, Lee T, Neveux LM, et al. Evaluation of cell-free fetal DNA as a second-trimester maternal serum marker of Down syndrome pregnancy. Clin Chem. 2003; 49(2):239-42.
  20. Lee T, LeShane ES, Messerlian GM, Canick JA, Farina A, Heber WW, et al. Down syndrome and cell-free fetal DNA in archived maternal serum. Am J Obstet Gynecol. 2002; 187(5):1217-21.
  21. Dar P, Curnow KJ, Gross SJ, Hall MP, Stosic M, Demko Z, et al. Clinical experience and follow-up with large scale single-nucleotide polymorphis-mebased noninvasive prenatal aneuploidy testing. Am J Obstet Gynecol. 2014; 211(5):527.e1-17.
  22. Rava RP, Srinivasan A, Sehnert AJ, Bianchi DW. Circulating fetal cell-free DNA fractions differ in autosomal aneuploidies and monosomy X. Clin Chem. 2014; 60(1):243-50.
  23. Wright A, Zhou Y, Weier JF, Caceres E, Kapidzic M, Tabata T, et al. Trisomy 21 is associated with variable defects in cytotrophoblast differentiation along the invasive pathway. Am J Med Genet A. 2004; 130(4):354-64.
  24. Ashoor G, Poon L, Syngelaki A, Mosimann B, Nicolaides KH. Fetal fraction in maternal plasma cell-free DNA at 11-13 weeks’ gestation: effect of maternal and fetal factors. Fetal Diagn Ther. 2012; 31(4):237-43.
  25. Ashoor G, Syngelaki A, Poon L, Rezende J, Nicolaides KJ. Fetal fraction in maternal plasma cell‐free DNA at 11–13 weeks' gestation: relation to maternal and fetal characteristics. Ultrasound Obstet Gynecol. 2013; 41(1):26-32.
  26. Haghiac M, Vora NL, Basu S, Johnson KL, Presley L, Bianchi DW, et al. Increased death of adipose cells, a path to release cell‐free DNA into systemic circulation of obese women. Obesity (Silver Spring). 2012; 20(11):2213-9.
  27. Al Nakib M, Desbriere R, Bonello N, Bretelle F, Boubli L, Gabert J, et al. Total and fetal cell-free DNA analysis in maternal blood as markers of placental insufficiency in intrauterine growth restriction. Fetal Diagn Ther. 2009; 26(1):24-8.
  28. Alberry MS, Maddocks DG, Hadi MA, Metawi H, Hunt LP, Abdel-Fattah SA, et al. Quantification of cell free fetal DNA in maternal plasma in normal pregnancies and in pregnancies with placental dysfunction. Am J Obstet Gynecol. 2009; 200(1):
    98.e1-6.
  29. Hui L. Noninvasive prenatal testing for aneuploidy using cell-free DNA–New implications for maternal health. Obstet Med. 2016; 9(4):148-52.
  30. Nitsche JF, Barnard A, Conrad S, Onslow M, Stephens N, Stringfellow J, et al. 29: The effect of maternal heparins and/or aspirin on the amount of cell-free fetal DNA in the maternal circulation. Am J Obstet Gynecol. 2017; 216(1):S20.
  31. Manokhina I, Singh TK, Robinson WP. Cell-free placental DNA in maternal plasma in relation to placental health and function. Fetal Diagn Ther. 2017; 41(4):258-64.
  32. Scott FP, Menezes M, Palma-Dias R, Nisbet D, Schluter P, da Silva Costa F, et al. Factors affecting cell-free DNA fetal fraction and the consequences for test accuracy. J Matern Fetal Neonatal Med. 2018; 31(14):1865-72.
  33. Nicolaides KH. Screening for fetal aneuploidies at 11 to 13 weeks. Prenat Diagn. 2011; 31(1):7-15.
  34. Nicolaides KH. Turning the pyramid of prenatal care. Fetal Diagn Ther. 2011; 29(3):183-96.
  35. Bredaki FE, Wright D, Akolekar R, Cruz G, Nicolaides KH. Maternal serum alpha-fetoprotein in normal pregnancy at 11–13 weeks’ gestation. Fetal Diagn Ther. 2011; 30(4):274-9.
  36. Kagan KO, Wright D, Spencer K, Molina F, Nicolaides KH. First‐trimester screening for trisomy 21 by free beta‐human chorionic gonadotropin and pregnancy‐associated plasma protein‐A: impact of maternal and pregnancy characteristics. Ultrasound Obstet Gynecol. 2008; 31(5):493-502.
  37. Pandya P, Wright D, Syngelaki A, Akolekar R, Nicolaides KH. Maternal serum placental growth factor in prospective screening for aneuploidies at 8-13 weeks’ gestation. Fetal Diagn Ther. 2012; 31(2):87-93.
  38. Metzenbauer M, Hafner E, Hoefinger D, Schuchter K, Stangl G, Ogris E, et al. Three-dimensional ultrasound measurement of the placental volume in early pregnancy: method and correlation with biochemical placenta parameters. Placenta. 2001; 22(6):602-5.
  39. Roland MC, Friis CM, Lorentzen B, Bollerslev J, Haugen G, Henriksen T. PP076. Gender differences in fetal growth and fetal-placental ratio in preeclamptic and normal pregnancies. Pregnancy Hypertens. 2013; 3(2):95.

 

Iranian Journal of Neonatology
Volume 12, Issue 1
Winter 2021
Pages 26-32