Validation of Highly Polymorphic Fluorescent Multiplex Short Tandem Repeat Systems Using Two Generations of DNA Sequencers

Validation studies are a crucial requirement before implementation of new genetic typing systems for clinical diagnostics or forensic identity. Two different fluorescence-based multiplex DNA profiling systems composed of amelogenin, HumD21S11 and HumFGA (referred to as multiplex 1A), and HumD3S1358,...

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Bibliographic Details
Published in:Journal of forensic sciences 1999-01, Vol.44 (1), p.133-166
Main Authors: Frégeau, CJ, Bowen, KL, Fourney, RM
Format: Article
Language:English
Subjects:
Animals
Blood Stains
Deoxyribonucleic acid
DNA
DNA, Bacterial - chemistry
DNA, Fungal - chemistry
Electrophoresis, Agar Gel
Female
Forensic sciences
Humans
Male
Polymorphism, Genetic
Polymorphism, Restriction Fragment Length
Primates
Sequence Analysis, DNA - methods
Sex Offenses
Tandem Repeat Sequences
Temperature
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Summary:Validation studies are a crucial requirement before implementation of new genetic typing systems for clinical diagnostics or forensic identity. Two different fluorescence-based multiplex DNA profiling systems composed of amelogenin, HumD21S11 and HumFGA (referred to as multiplex 1A), and HumD3S1358, HumD21S11 and HumFGA (multiplex 1B) have been evaluated for use in forensic identification using the Applied Biosystems Model 373A and Prism™ 377 DNA Sequencers, respectively. Experiments were aimed at defining the limit of target DNA required for reliable profiling, the level of degradation that would still permit amplification of the short tandem repeat (STR) loci examined, and the robustness of each locus in the multiplexes after samples were exposed to environmental insults. In addition, the specificity of the multiplexes was demonstrated using nonhuman DNAs. Forensically relevant samples such as cigarette butts, chewing gum, fingernails and envelope flaps were processed using both an organic extraction procedure and a QIAamp protocol. DNAs and resultant multiplex STR profiles were compared. The validation of the triplex STR systems was extended to include over 140 nonprobative casework specimens and was followed with a close monitoring of initial casework (over 300 exhibits). Our results document the robustness of these multiplex STR profiling systems which, when combined with other multiplex systems, could provide a power of discrimination of approximately 0.9999.
ISSN:0022-1198
1556-4029
DOI:10.1520/JFS14425J