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<p>Forensic DNA analysis is essential in criminal investigations, with blood being a primary biological sample. However, blood samples are often subjected to prolonged exposure to environmental conditions, leading to degradation and compromising DNA recovery. This study evaluates the effectiveness of polyethyleneimine functionalized magnetic iron oxide nanoparticle (PEI-IONP)-based DNA isolation method with the traditional phenol–chloroform approach and the Qiagen kit-based method. Blood samples obtained from 10 healthy volunteers were exposed to different time intervals—0&#xa0;h (fresh sample), 24&#xa0;h, 72&#xa0;h, and 1&#xa0;week. DNA was isolated from each time point sample using the three methods, and the DNA concentration, purity, and integrity were assessed. The PEI-IONP-based method consistently yielded significantly higher mean DNA concentrations compared to the other two methods at all time points. At 0&#xa0;h, the PEI-IONP-based method yielded a mean DNA concentration of 33.5 ± 2.6&#xa0;ng/µL, outperforming the Qiagen kit (28.0 ± 3.8&#xa0;ng/µL) and phenol–chloroform (25.25 ± 4.3&#xa0;ng/µL) methods. Even after 1&#xa0;week, the PEI-IONP-based method maintained significantly higher mean DNA concentration (23.75 ± 5&#xa0;ng/µL) compared to the Qiagen kit-based (13.5 ± 4.8&#xa0;ng/µL) and phenol–chloroform (9.0 ± 5.2&#xa0;ng/µL) methods. Mean DNA purity ratios remained stable for the PEI-IONP-based method (1.82–1.86), whereas the other methods showed declining purity. Agarose gel electrophoresis revealed less smearing with the PEI-IONP-based approach compared to the other two methods at later time points. These findings suggest that the PEI-IONP-based approach is a robust and effective alternative for forensic applications, where sample integrity and delayed processing are significant concerns.</p>

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Comparative Evaluation of Polyethyleneimine Functionalized Magnetic Iron-Oxide Nanoparticle-Based Method, Phenol–Chloroform Method, and Qiagen Kit-Based Method for Isolation of Deoxyribonucleic Acid (DNA) from Blood Samples at Different Time Points

  • 期刊论文

  • 发表日期：

    2025年5月19日

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  • Imran Khan,
  • Gaurav Kaushik,
  • Chaitenya Verma,
  • Richa Vashishtha,
  • Nawaid Hussain Khan,
  • Vinay Kumar

摘要

Forensic DNA analysis is essential in criminal investigations, with blood being a primary biological sample. However, blood samples are often subjected to prolonged exposure to environmental conditions, leading to degradation and compromising DNA recovery. This study evaluates the effectiveness of polyethyleneimine functionalized magnetic iron oxide nanoparticle (PEI-IONP)-based DNA isolation method with the traditional phenol–chloroform approach and the Qiagen kit-based method. Blood samples obtained from 10 healthy volunteers were exposed to different time intervals—0 h (fresh sample), 24 h, 72 h, and 1 week. DNA was isolated from each time point sample using the three methods, and the DNA concentration, purity, and integrity were assessed. The PEI-IONP-based method consistently yielded significantly higher mean DNA concentrations compared to the other two methods at all time points. At 0 h, the PEI-IONP-based method yielded a mean DNA concentration of 33.5 ± 2.6 ng/µL, outperforming the Qiagen kit (28.0 ± 3.8 ng/µL) and phenol–chloroform (25.25 ± 4.3 ng/µL) methods. Even after 1 week, the PEI-IONP-based method maintained significantly higher mean DNA concentration (23.75 ± 5 ng/µL) compared to the Qiagen kit-based (13.5 ± 4.8 ng/µL) and phenol–chloroform (9.0 ± 5.2 ng/µL) methods. Mean DNA purity ratios remained stable for the PEI-IONP-based method (1.82–1.86), whereas the other methods showed declining purity. Agarose gel electrophoresis revealed less smearing with the PEI-IONP-based approach compared to the other two methods at later time points. These findings suggest that the PEI-IONP-based approach is a robust and effective alternative for forensic applications, where sample integrity and delayed processing are significant concerns.