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Original Research article

Optimization of Pre-treatment Conditions for Direct PCR Analysis of Human  Bloodstains at Various Storage Durations

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Wimbuh Tri Widodo — Master of Forensic Science, Postgraduate School, Universitas Airlangga, Surabaya, 60286, Indonesia
Sonny Kristianto — Master of Forensic Science, Postgraduate School, Universitas Airlangga, Surabaya, 60286, Indonesia
Rury Eryna Putri — Master of Forensic Science, Postgraduate School, Universitas Airlangga, Surabaya, 60286, Indonesia
Ahmad Yudianto — Human Genetics Laboratory, Institute of Tropical Disease, Universitas Airlangga, Surabaya, 60115, Indonesia
Fatimah — Biotechnology Study Program, Postgraduate School, Gadjah Mada University, Yogyakarta, 55281, Indonesia
Lalu Unsunnidhal — Laboratory of Molecular Biotechnology, Graduate School of Bioagricultural Sciences, Nagoya University, 464-8601, Nagoya, Japan
Published 15 Apr 2026 Vol. 43 No. 1 (2026), pp. 32-36
❝ How to cite 🔗 https://doi.org/10.48165/jfmt.2026.43.01.11
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Abstract

The polymerase chain reaction (PCR) has undergone significant advancements and has been applied in various fields,  including forensics. In the field of forensic analysis, rapidity and efficiency are paramount. One promising technique  is direct PCR, which involves conducting PCR analysis without DNA extraction, facilitating a more rapid and efficient  analysis. However, treating samples used directly as templates is essential to ensure the PCR reaction can proceed. This  study involved optimizing the treatment of blood samples (incubation at 60°C and 90°C for 10 minutes) prior to their  use as a direct PCR template and evaluating the effectiveness of direct PCR on human bloodstains incubated for various  durations (0, 7, 14, 21, and 30 days) using primers of the human cytochrome b gene. The gene has significant variation  between species, but limited variation within the species. The results indicated that the cytochrome b gene primers suc cessfully amplified human DNA. Samples incubated at 90°C for 10 minutes or at 60°C for 10 minutes could be used as  direct PCR templates. Human bloodstains remained analyzable using direct PCR up to 30 days following the initial sampling. This study provides valuable insights for forensic experts, enabling them to expedite and enhance the efficiency of  the PCR process in forensic applications. 

 

KeywordsDNADirect PCRCrimeCytochrome bBlood

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Copyright © 2026 Wimbuh Tri Widodo, Sonny Kristianto, Rury Eryna Putri, Ahmad Yudianto, Fatimah, Lalu Unsunnidhal (Author)