Abstract
A quadruplex reaction has been developed which amplifies the short tandem repeat (STR) loci HUMVWA31/A, HUMTHO1, HUMF13A1 and HUMFES/FPS. Detection of the PCR products employs denaturing poly-acrylamide gels coupled with fluorescent-based technology. This system has been evaluated for use in routine forensic casework and has been shown to be both robust and reproducible. The quadruplex reaction is as sensitive as the commercially available HLA DQα Amplitype typing system and can be used on both degraded and aged material. The problems of environmental contamination have been shown to be limited provided strict procedural practices are followed — i.e. physical separation of sample extraction and amplified products; the use of dedicated equipment such as pipettes; the separation of amplification preparation area. The ability of the system to detect mixtures and the successful analysis of case stains has shown that this system is well suited as a tool for forensic investigation.
Zusammenfassung
Eine Quadruplex-Reaktion wurde entwickelt, die die Short tandem repeat loci (STR) HUMVWA31/1, HUMTHO1, HUMF13A1 und HUMFES/FPS amplifiziert. Zum Nachweis der PCR-Produkte werden denaturierende Polyacrylamid-Gele benutzt, in Kombination mit einer Fluoreszenz-basierten Technologie. Dieses System wurde fÜr den Gebrauch in der routinemäßigen Fallbearbeitung evaluiert und hat sich als robust und reproduzierbar erwiesen. Die Quadruplex-Reaktion ist genau so empfindlich wie das kommerzielle HLADQα-Ampli-type Typisierungssystem und kann sowohl an degradiertem als auch an gealtertem Material verwendet werden. Die Probleme einer umgebungsbedingten Kontamination haben sich als begrenzt erwiesen, vorausgesetzt, daß strikte prozedurale Praktiken benutzt werden, d. h. physische Trennung der Extraktion und der amplifizierten Produkte-, Benutzung persönlicher Einrichtungen, wie Pipetten; Abtrennung des Bereichs der Amplifikationsvorbereitung; Benutzung der Laminar Flow-Einrichtung für die Vorbereitung der Amplifikation. Die FÄhigkeit des Systems, Mischungen zu erkennen und die erfolgreiche Analyse von Spurenfällen haben gezeigt, daß dieses System gut geeignet ist als ein Werkzeug für forensische Untersuchung.
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Lygo, J.E., Johnson, P.E., Holdaway, D.J. et al. The validation of short tandem repeat (STR) loci for use in forensic casework. Int J Leg Med 107, 77–89 (1994). https://doi.org/10.1007/BF01225493
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DOI: https://doi.org/10.1007/BF01225493