Порівняльна ефективність наборів PrepFiler™ Forensic DNA Extraction Kit  та NucleoSpin® DNA Forensic у контексті виділення ДНК з проблемних зразків для STR-аналізу

L. I. Kucherenko; D. V. Zubryk; I. V. Pavliuk; H. R. Nimenko

doi:10.14739/2409-2932.2025.3.332974

Authors

L. I. Kucherenko Zaporizhzhia State Medical and Pharmaceutical University, Ukraine https://orcid.org/0000-0003-2229-0232
D. V. Zubryk Zaporizhzhia Research Experimental Forensic Center of the Ministry of Internal Affairs of Ukraine, Ukraine https://orcid.org/0009-0004-2619-1463
I. V. Pavliuk Zaporizhzhia Research Experimental Forensic Center of the Ministry of Internal Affairs of Ukraine, Ukraine https://orcid.org/0000-0002-6423-8777
H. R. Nimenko Zaporizhzhia State Medical and Pharmaceutical University, Ukraine https://orcid.org/0000-0002-6486-5113

DOI:

https://doi.org/10.14739/2409-2932.2025.3.332974

Keywords:

molecular genetics, DNA, analysis, identification, genetic locus

Abstract

In modern forensic practice, molecular genetic studies play an extremely important role, especially in wartime. DNA identification allows you to identify a person by traces of biological origin, which has become critically important for the identification of the dead and missing. The effectiveness of such studies largely depends on the choice of the optimal method of DNA extraction, which ensures the reliability and speed of results. In this regard, the choice of the most reliable, sensitive and resource-efficient method of DNA extraction becomes not only a technical but also a strategic task that directly affects the success of investigations and the reduction of the number of missing persons, which is extremely important in the humanitarian dimension of modern war.

Aim. Comparative analysis of two commercial kits for manual DNA extraction — PrepFiler™ Forensic DNA Extraction Kit (Applied Biosystems) and NucleoSpin® DNA Forensic (Macherey-Nagel) — to determine the feasibility of their use for different types of biological samples and traces and categories of expert workload.

Materials and methods. The study was conducted on the basis of the analysis of samples of trace biological material, in particular blood stains, saliva, tissue fragments and traces from the surfaces of objects. Two sets of reagents were used for DNA extraction: PrepFiler™ Forensic DNA Extraction Kit (Thermo Fisher Scientific) and NucleoSpin® DNA Forensic (Macherey-Nagel), 100 reactions each. DNA extraction was carried out according to the protocols provided by the manufacturers. The quality and quantity of the isolated DNA were determined by real-time polymerase chain reaction using the Applied Biosystems 7300 Real-Time PCR System. Further genotyping was performed by capillary electrophoresis on the Applied Biosystems 3500 Genetic Analyzer.

Results. Comparative analysis showed that NucleoSpin® DNA Forensic is optimal for fast and convenient DNA isolation from simple reference samples such as saliva or blood, where no significant degradation or inhibitors are observed. It provides sufficient DNA concentration in a short time, however, in cases of working with complex or degraded samples (especially bone remains), partial STR profiles were more often observed. In contrast, PrepFiler™ and, in particular, PrepFiler™ BTA demonstrated better performance under such difficult conditions, providing complete profiles even in the presence of inhibitors or prolonged degradation. This makes it a reasonable choice for forensic cases with problematic samples, despite the higher cost.

Conclusions. Molecular genetic research is a key tool in modern forensic practice, in particular for identifying a person from biological samples. The use of DNA identification methods based on polymerase chain reaction (PCR) ensures high accuracy and reliability of forensic examinations in criminal and civil cases, and also plays an important role in the identification of the deceased and missing, especially in wartime conditions. A comparative analysis of two commercial kits for manual DNA extraction — PrepFiler™ Forensic DNA Extraction Kit and NucleoSpin® DNA Forensic — showed that both kits have their advantages and can be effectively used depending on the type of biological material and the conditions of the examination. The choice of the optimal DNA extraction method significantly affects the quality and quantity of the obtained genetic material, which, in turn, determines the reliability of DNA profiling and the speed of the expert laboratory.

Author Biographies

L. I. Kucherenko, Zaporizhzhia State Medical and Pharmaceutical University

PhD, DSc, Professor, Head of the Department of Pharmaceutical Organic and Bioorganic Chemistry

D. V. Zubryk, Zaporizhzhia Research Experimental Forensic Center of the Ministry of Internal Affairs of Ukraine

Head of the Department of Biological Research and Accounting

I. V. Pavliuk, Zaporizhzhia Research Experimental Forensic Center of the Ministry of Internal Affairs of Ukraine

PhD, Senior Judicial Expert in the Sector for the Study of Narcotic Drugs, Psychotropic Substances, their Analogues and Precursors in the Materials and Products Research Division

H. R. Nimenko, Zaporizhzhia State Medical and Pharmaceutical University

PhD, Associate Professor of the Department of Pharmaceutical, Organic and Bioorganic Chemistry

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