Enhancing trace DNA profile recovery in forensic casework using the amplicon RX post-PCR clean-up kit

Abstract This study evaluated the effectiveness of the amplicon RX post-PCR clean-up kit in enhancing trace DNA profile recovery from forensic casework samples amplified using the GlobalFiler PCR amplification kit. The impact of post-PCR clean-up on allele recovery and signal intensity was assessed...

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Main Authors: Naeema S. Aljanahi, Salem K. Alketbi, Maryam M. Almheiri, Suaad A. Alshehhi, Afra N. Sanqoor, Hussein J. Alghanim
Format: Article
Language:English
Published: Nature Portfolio 2025-01-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-88164-4
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author Naeema S. Aljanahi
Salem K. Alketbi
Maryam M. Almheiri
Suaad A. Alshehhi
Afra N. Sanqoor
Hussein J. Alghanim
author_facet Naeema S. Aljanahi
Salem K. Alketbi
Maryam M. Almheiri
Suaad A. Alshehhi
Afra N. Sanqoor
Hussein J. Alghanim
author_sort Naeema S. Aljanahi
collection DOAJ
description Abstract This study evaluated the effectiveness of the amplicon RX post-PCR clean-up kit in enhancing trace DNA profile recovery from forensic casework samples amplified using the GlobalFiler PCR amplification kit. The impact of post-PCR clean-up on allele recovery and signal intensity was assessed in both trace casework samples and control samples across a range of DNA concentrations. The results showed that the amplicon RX method significantly improved allele recovery compared to the 29-cycle protocol (p = 8.30 × 10−12) and achieved slightly better results than the 30-cycle protocol (p = 0.019). Additionally, the Amplicon RX method demonstrated a significant increase in signal intensity (p = 2.70 × 10−4), reflecting improved sensitivity in detecting trace DNA profiles compared to the 30-cycle protocol. In the evaluation of control samples, the amplicon RX method consistently outperformed both the 29- and 30-cycle protocols, especially at lower DNA concentrations (D3: 0.001 ng/µL). While the performance of all methods declined at the lowest concentration (D4: 0.0001 ng/µL), the Amplicon RX method still demonstrated superior allele recovery (p = 0.014 compared to 29 cycles; p = 0.011 compared to 30 cycles). Therefore, the Amplicon RX method should be widely adopted in forensic laboratories to enhance the analysis of extremely low-template and compromised samples. These findings highlight the potential of the amplicon RX post-PCR clean-up kit to improve trace DNA analysis in forensic casework. Further research is recommended to validate these results and explore its broader application in forensic DNA analysis, particularly in complex DNA mixtures and extremely low-template samples.
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spelling doaj-art-a8b2a714938a43eab3487abaf2aebd842025-02-02T12:19:25ZengNature PortfolioScientific Reports2045-23222025-01-0115111010.1038/s41598-025-88164-4Enhancing trace DNA profile recovery in forensic casework using the amplicon RX post-PCR clean-up kitNaeema S. Aljanahi0Salem K. Alketbi1Maryam M. Almheiri2Suaad A. Alshehhi3Afra N. Sanqoor4Hussein J. Alghanim5The Biology and DNA Section, General Department of Forensic Science and Criminology, Dubai Police General Head QuartersThe Biology and DNA Section, General Department of Forensic Science and Criminology, Dubai Police General Head QuartersThe Biology and DNA Section, General Department of Forensic Science and Criminology, Dubai Police General Head QuartersThe Biology and DNA Section, General Department of Forensic Science and Criminology, Dubai Police General Head QuartersThe Biology and DNA Section, General Department of Forensic Science and Criminology, Dubai Police General Head QuartersThe Biology and DNA Section, General Department of Forensic Science and Criminology, Dubai Police General Head QuartersAbstract This study evaluated the effectiveness of the amplicon RX post-PCR clean-up kit in enhancing trace DNA profile recovery from forensic casework samples amplified using the GlobalFiler PCR amplification kit. The impact of post-PCR clean-up on allele recovery and signal intensity was assessed in both trace casework samples and control samples across a range of DNA concentrations. The results showed that the amplicon RX method significantly improved allele recovery compared to the 29-cycle protocol (p = 8.30 × 10−12) and achieved slightly better results than the 30-cycle protocol (p = 0.019). Additionally, the Amplicon RX method demonstrated a significant increase in signal intensity (p = 2.70 × 10−4), reflecting improved sensitivity in detecting trace DNA profiles compared to the 30-cycle protocol. In the evaluation of control samples, the amplicon RX method consistently outperformed both the 29- and 30-cycle protocols, especially at lower DNA concentrations (D3: 0.001 ng/µL). While the performance of all methods declined at the lowest concentration (D4: 0.0001 ng/µL), the Amplicon RX method still demonstrated superior allele recovery (p = 0.014 compared to 29 cycles; p = 0.011 compared to 30 cycles). Therefore, the Amplicon RX method should be widely adopted in forensic laboratories to enhance the analysis of extremely low-template and compromised samples. These findings highlight the potential of the amplicon RX post-PCR clean-up kit to improve trace DNA analysis in forensic casework. Further research is recommended to validate these results and explore its broader application in forensic DNA analysis, particularly in complex DNA mixtures and extremely low-template samples.https://doi.org/10.1038/s41598-025-88164-4Forensic scienceTrace DNATouch DNADNA recoveryGlobalFiler PCR amplification kitAmplicon RX post-PCR clean-up kit
spellingShingle Naeema S. Aljanahi
Salem K. Alketbi
Maryam M. Almheiri
Suaad A. Alshehhi
Afra N. Sanqoor
Hussein J. Alghanim
Enhancing trace DNA profile recovery in forensic casework using the amplicon RX post-PCR clean-up kit
Scientific Reports
Forensic science
Trace DNA
Touch DNA
DNA recovery
GlobalFiler PCR amplification kit
Amplicon RX post-PCR clean-up kit
title Enhancing trace DNA profile recovery in forensic casework using the amplicon RX post-PCR clean-up kit
title_full Enhancing trace DNA profile recovery in forensic casework using the amplicon RX post-PCR clean-up kit
title_fullStr Enhancing trace DNA profile recovery in forensic casework using the amplicon RX post-PCR clean-up kit
title_full_unstemmed Enhancing trace DNA profile recovery in forensic casework using the amplicon RX post-PCR clean-up kit
title_short Enhancing trace DNA profile recovery in forensic casework using the amplicon RX post-PCR clean-up kit
title_sort enhancing trace dna profile recovery in forensic casework using the amplicon rx post pcr clean up kit
topic Forensic science
Trace DNA
Touch DNA
DNA recovery
GlobalFiler PCR amplification kit
Amplicon RX post-PCR clean-up kit
url https://doi.org/10.1038/s41598-025-88164-4
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