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Croatian Medical Journal, Vol. 58 No. 1, 2017.

Original scientific paper

https://doi.org/10.3325/cmj.2017.58.26

PowerPlex® Fusion 6C System: evaluation study for analysis of casework and database samples

Selena Cisana ; University of Brescia - Department of Medical and Surgical Specialties Radiological Sciences and Public Health - Forensic Medicine Unit, Brescia, Italy
Nicoletta Cerri ; University of Brescia - Department of Medical and Surgical Specialties Radiological Sciences and Public Health - Forensic Medicine Unit, Brescia, Italy
Alessandro Bosetti ; Promega Italia s.r.l. Via Decembrio 28, 20137 Milano, Italy
Andrea Verzeletti ; University of Brescia - Department of Medical and Surgical Specialties Radiological Sciences and Public Health - Forensic Medicine Unit, Brescia, Italy
Venusia Cortellini ; University of Brescia - Department of Medical and Surgical Specialties Radiological Sciences and Public Health - Forensic Medicine Unit, Brescia, Italy

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Abstract

Aim To report on the successful analysis of amplicons obtained with PowerPlex® Fusion 6C System, a highly robust 27-plex genotyping kit developed for human identification laboratories, on the Applied Biosystems® 3500 Genetic Analyzer.
Method We performed characterization and evaluation studies following the Scientific Working Group on DNA Analysis Methods (SWGDAM) validation guidelines, examining several critical areas of kit performance. We report the results of sensitivity, robustness, heterozygous peak height ratio, precision, concordance, caseworks, and mixture interpretations. We tested sensitivity, using serial dilutions of control DNA.
Results The minimum amount of input DNA resulting in a full profile was 125 pg. Inhibition, inducted by urea, showsed a progressively fragmentation of DNA and a full profile was obtained until 1M of inhibitor factor. To test the profile quality, casework samples were extracted with different extraction methods: Chelex®100, QIAmp DNA Micro Kit and Phenol-Chloroform extraction. The results demonstrated that extraction chemistries do not have affect on amplification performance. Concordance check was performed by typing some casework samples and comparing the typing results with those obtained with other available kits. Thus, concordance was expected and supported by the data.
Conclusion Reliable DNA typing results can be obtained using this new kit, demonstrating its effectiveness and utility in forensic analysis.

Keywords

Hrčak ID:

181403

URI

https://hrcak.srce.hr/181403

Publication date:

15.2.2017.

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