A Novel Nanoparticle-Based Method to Isolate DNA From Dried Saliva and Semen Samples

Document Type: Original Articles

Authors

Department of Life Sciences, School of Sciences, Gujarat University, Ahmedabad, India.

Abstract

Objectives: Saliva and semen stains may be found at crime scene (suicide, homicide, kidnapping, rape, sexual assault, poisoning, etc.). Through, which DNA can be isolated and individual can be identified. DNA can be of great importance in forensic science investigation to identify individuals. Currently, there are enormous methods to isolate DNA from biological fluids. The current study employed nanoparticle–biomolecule conjugate method to isolate DNA. Materials & Methods: A nanoparticle-biomolecule conjugate is a magnetic nanoparticle with biomolecules attached to its surface. Saliva and semen were collected using cotton swabs from different surfaces (tile, steel, aluminum, glass, iron, wood, etc.) on which saliva and semen were poured and allowed to dry earlier as to create simulated crime scene. Magnetic nanoparticles (Fe3O4) act as solid phase support to isolate DNA from saliva and semen stains. In the current study, cells and spermatozoa were extracted from saliva and semen swabs, respectively. Cell lysis was performed to extract epithelial cells from saliva and spermatozoa from semen and magnetic nanoparticles and biomolecule conjugate were added to the lysed cells to bind to DNA. Results: Application of external magnetic field helped to settle down nanoparticle-biomolecule conjugate and thus separate DNA from other cell components. Spectrophotometer Nanodrop technology was employed to perform the readings, which confirmed the presence of DNA at 260/280 ratio and quality of DNA was compared with the DNA isolated using the two different methods.Conclusion: The current study aimed at developing all new methods to isolate DNA from dried saliva and semen samples using magnetic nanoparticles by reviewing the methods developed earlier to isolate DNA from blood and tissue using magnetic nanoparticles. With modifications in the method and chemical preparation, a new method was successfully developed. DNA was isolated successfully; and confirmation was made by analyzing DNA through Nanodrop spectrophotometer.

Keywords


  1. Sun F, Reichenberger EJ. Saliva as a source of genomic DNA for genetic studies: Review of current methods and applications. Journal of Oral Health and Dental Management. 2014; 13(2):217-22. [PMID]
  2. Nunes LAS, Brenzikofer R, Macedo DV. Reference intervals for saliva analytes collected by a standardized method in a physically active population. Clinical Biochemistry, 44(17-18):1440-4. [DOI:10.1016/j.clinbiochem.2011.09.012] [PMID]
  3. Mapes AA, Kloosterman AD, de Poot CJ. DNA in the criminal justice system: The DNA success story in perspective. Journal of forensic sciences. 2015; 60(4):851-6. [DOI:10.1111/1556-4029.12779] [PMID]
  4. Butler JM. Forensic DNA typing: Biology, technology and genetics of STR markers. Amsterdam: Elsevier Science; 2005.
  5. Johnson D, Peterson J, Sommers I, Baskin D. Use of forensic science in investigating crimes of sexual violence: Contrasting its theoretical potential with empirical realities. Violence Against Women. 2012; 18(2):193-222. [DOI:10.1177/1077801212440157] [PMID]
  6. Lee HC, Ladd C. Preservation and collection of biological evidence. Croatian Medical Journal. 2001; 42(3):225-8. [PMID]
  7. Raymond JJ, van Oorschot RA, Gunn PR, Walsh SJ, Roux C. Trace evidence characteristics of DNA: A preliminary investigation of the persistence of DNA at crime scenes. Forensic Science International: Genetics. 2009; 4(1):26-33. [DOI:10.1016/j.fsigen.2009.04.002] [PMID]
  8. Bozzo WR, Colussi AG, Ortiz MI, Lojo MM. DNA recovery from different evidences in 300 cases of sexual assault. Forensic Science International: Genetics Supplement Series. 2009; 2(1):141-2. [DOI:10.1016/j.fsigss.2009.08.185]
  9. Khaldi N, Miras A, Botti K, Benali L, Gromb S. Evaluation of three rapid detection methods for the forensic identification of seminal fluid in rape cases. Journal of Forensic Science. 2004; 49(4):1-5. [DOI:10.1520/JFS2003317] [PMID]
  10. Magalhães T, Dinis Oliveira RJ, Silva B, Corte Real F, Nuno Vieira D. Biological evidence management for DNA analysis in cases of sexual assault. The Scientific World Journal. 2015; 2015:365674. [DOI:10.1155/2015/365674] [PMID] [PMCID]
  11. Newton M. The forensic aspects of sexual violence. Best Practice & Research Clinical Obstetrics & Gynaecology. 2013; 27(1):77-90. [DOI:10.1016/j.bpobgyn.2012.08.020] [PMID]
  12. Stangegaard M, Hjort BB, Hansen TN, Hoflund A, Mogensen, HS Hansen AJ, et al. Automated extraction of DNA from biological stains on fabric from crime cases: A comparison of a manual and three automated methods. Forensic Science International: Genetics. 2013; 7(3):384-8. [DOI:10.1016/j.fsigen.2012.12.009] [PMID]
  13. Prodělalova J, Rittich B, Španova A, Petrova K, Beneš MJ. Isolation of genomic DNA using magnetic cobalt ferrite and silica particles. Journal of Chromatography A. 2004; 1056(1-2):43-8. [DOI:10.1016/S0021-9673(04)01448-7]
  14. Saiyed ZM, Bochiwal C, Gorasia H, Telang SD, Ramchand CN. Application of magnetic particles (Fe3O4) for isolation of genomic DNA from mammalian cells. Analytical Biochemistry. 2006; 356(2):306-8. [DOI:10.1016/j.ab.2006.06.027] [PMID]
  15. John SW, Weitzner G, Rozen R, Scriver CR. A rapid procedure for extracting genomic DNA from leukocytes. Nucleic Acids Research. 1991; 19(2):408. [DOI:10.1093/nar/19.2.408] [PMID] [PMCID]
  16. Saiyed ZM, Ramchand CN, Telang SD. Isolation of genomic DNA using magnetic nanoparticles as a solid-phase support. Journal of Physics: Condensed Matter. 2008; 20(20):204153. [DOI:10.1088/0953-8984/20/20/204153] [PMID]
  17. Safarik I, Safarikova M. Magnetic nanoparticles and biosciences. Monatshefte für Chemie-Chemical Monthly. 2002; 133(6):737-59. [DOI:10.1007/s007060200047]