Journal of Assisted Reproduction and Genetics

, Volume 21, Issue 11, pp 397–400 | Cite as

Modified Isocratic Capillary Electrophoresis Detection of Cell-free DNA in Semen

  • Jennifer S. Chou
  • John D. Jacobson
  • William C. Patton
  • Alan King
  • Philip J. Chan

Abstract

Purpose: The objectives were: i) to analyze semen for the presence of cell-free DNA and ii) to determine the association between sperm parameters and cell-free DNA.

Methods: Cell-free DNA in semen (N=25 cases) were detected using the modified capillary gel electrophoresis (CE) procedure. SYBR-Gold was used to stain high (12 Kb) and low (1 Kb) molecular weight DNA fragments and the images analyzed.

Results: The quantity of low-molecular weight cell-free DNA was positively correlated to rapid progression, curvilinear velocity (>40 μ/s), normal strict morphology and capacitation index. High-molecular weight cell-free DNA intensity index was negatively correlated to post-wash hyperactivation. Sperm concentration was not related to cell-free DNA quantity. The sperm freezing process did not increase cell-free DNA but reduced the more labile low-molecular weight cell-free DNA.

Conclusions: Cell-free DNA present in semen was correlated to important sperm parameters linked to normal sperm function. The data suggested the possible use of cell-free DNA as a marker of semen quality. This study reports on the novel finding of cell-free DNA released along with sperm during each ejaculation.

Apoptosis and active secretion circulating nucleic acids infertility spermatozoa 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Lo YM, Corbetta N, Chamberlain PF, Rai V, Sargent IL, Redman CW, Wainscoat JS: Presence of fetal DNA in maternal plasma and serum. Lancet 1997; 16: 485–487Google Scholar
  2. 2.
    Lo YM, Tein MS, Lau TK, Haines CJ, Leung TN, Poon PM, Wainscoat JS, Johnson PJ, Chang AM, Hjelm NM: Quantitative analysis of fetal DNA in maternal plasma and serum: Implications for noninvasive prenatal diagnosis. Am J Hum Genet 1998; 62: 768–775Google Scholar
  3. 3.
    Mandel P, Métais P: Les acides nucléiques du plasma sanguin chez l’homme. C.R. Acad Sci Paris 1948; 142: 241–243Google Scholar
  4. 4.
    Tan EM, Schur PH, Carr RI, Kunkel HG: Deoxybonucleic acid (DNA) and antibodies to DNA in the serum of patients with systemic lupus erythematosus. J Clin Invest 1966; 45: 1732–1740Google Scholar
  5. 5.
    Leon SA, Shapiro B, Sklaroff DM, Yaros MJ: Free DNA in the serum of cancer patients and the effect of therapy. Cancer Res 1977; 37: 646–650Google Scholar
  6. 6.
    Tsuda T, Nakagawa G, Sato M, Yagi K: Separation of nucleotides by high-voltage capillary electrophoresis. J Appl Biochem 1983; 5: 330–336Google Scholar
  7. 7.
    Kasper TJ, Melera M, Gozel P, Brownlee RG: Separation and detection of DNA by capillary electrophoresis. J Chromatogr 1988; 23: 303–312Google Scholar
  8. 8.
    Cohen AS, Najarian DR, Paulus A, Guttman A, Smith JA, Karger BL. Rapid separation and purification of oligonucleotides by high-performance capillary gel electrophoresis. Proc Natl Acad Sci U S A 1988; 85: 9660–9663Google Scholar
  9. 9.
    Checa ML, Dunner S, Canon J: Prediction of X and Y chromosome content in bovine sperm by using DNA pools through capillary electrophoresis. Theriogenology 2002; 58: 1579–1586Google Scholar
  10. 10.
    Chan PJ, Corselli JU, Patton WC, Jacobson JD, King A: Enhanced fertility after heat-induced hyperactivation. Fertil Steril 1998; 69: 118–121Google Scholar
  11. 11.
    Rojas FJ, La AT, Ord T, Patrizio P, Balmaceda JP, Silber SJ, Asch RH: Penetration of zona-free hamster oocytes using human sperm aspirated from the epididymis of men with congenital absence of the vas deferens: Comparison with human in vitro fertilization. Fertil Steril 1992; 58: 1000–1005Google Scholar
  12. 12.
    Perez SM, Chan PJ, Patton WC, King A: Silane-coated silica particle colloid processing of human sperm. J Assist Reprod Genet 1997; 14: 388–393Google Scholar
  13. 13.
    Giacona MB, Ruben GC, Iczkowski KA, Roos TB, Porter DM, Sorenson GD: Cell-free DNA in human blood plasma: Length measurements in patients with pancreatic cancer and healthy controls. Pancreas 1998; 17: 89–97Google Scholar
  14. 14.
    Wu TL, Zhang D, Chia JH, Tsao KH, Sun CF, Wu JT: Cell-free DNA: Measurement in various carcinomas and establishment of normal reference range. Clin Chim Acta 2002; 321: 77–87Google Scholar
  15. 15.
    Rogers JC, Boldt D, Kornfeld S, Skinner A, Valeri CR: Excretion of deoxyribonucleic acid by lymphocytes stimulated with phytohemagglutinin or antigen. Proc Natl Acad Sci USA 1972; 69: 1685–1689Google Scholar
  16. 16.
    Holmgren L, Szeles A, Rajnavolgyi E, Folkman J, Klein G, Ernberg I, Falk KI: Horizontal transfer of DNA by the uptake of apoptotic bodies. Blood 1999; 93: 3956–3963Google Scholar
  17. 17.
    Chomont N, Gresenguet G, Levy M, Hocini H, Becquart P, Matta M, Tranchot-Diallo J, Andreoletti L, Carreno MP, Kazatchkine MD, Belec L: Detection of Y chromosome DNA as evidence of semen in cervicovaginal secretions of sexually active women. Clin Diagn Lab Immunol. 2001; 8: 955–958Google Scholar

Copyright information

© Springer Science+Business Media, Inc. 2004

Authors and Affiliations

  • Jennifer S. Chou
    • 1
  • John D. Jacobson
    • 1
  • William C. Patton
    • 1
  • Alan King
    • 1
  • Philip J. Chan
    • 1
  1. 1.Department of Gynecology and ObstetricsLoma Linda University School of MedicineLoma LindaU.S.A.

Personalised recommendations