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The AluQuant™ (Promega Corporation) liquid hybridization DNA quantitation method was evaluated on an automated robotic platform (Biomek® 2000, Beckman Coulter, Fullerton, CA) for use in forensic PCR-STR systems. DNA from bloodstains and buccal swabs was extracted by three different methods: Chelex, Qiagen™ and DNA IQ (Promega). Samples were quantitated using both the Quantiblot and the AluQuant™ systems.Concordance between methodswas determined by comparing the average AluQuant™ DNA concentrations for samples having matching (binned) Quantiblot values. Studies testing the “accuracy” (STR analysis), precision, sensitivity, and specifies specificity of the AluQuant™ method were also conducted. The effect of inhibitors (carpet, denim, and suede) was evaluated. The results indicate that the AluQuant™ quantitation system equals the Quantiblot system in “accuracy”, sensitivity, precision, and primate-specificity. While extracts from denim and suede affected (inhibited) both systems minimally, the carpet extracts produced a sharp increase in DNA quantitation values in the AluQuant™ but not the Quantiblot system. The speed and user-friendliness of the AluQuant™ system on a robotic platform offer specific advantages to the forensic community. |
