Detection sensibility and amplification threshold establishment: Internal validation of Investigator® Quantiplex® PRO in a genetic forensic laboratory in central Brazil
- Forensic Research & Criminology International Journal
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Advaldo Carlos Souza Neto, Kamilla Nogueira Pimentel, Laryssa Silva de Andrade
Abstract
The present study aims to describe the internal validation of DNA quantification process in the Forensic Biology and DNA Laboratory (LBDF) of Goiás, Brazil. The sensitivity, precision and accuracy of the Investigator® Quantiplex® Pro, a real-time human DNA quantification kit, were evaluated. We used three blood samples with known genetic profile and tested the parameters in triplicate with DNA concentration ranging from 1ng/ µl to 0.0005ng/µl. We observed that the kit has high sensibility, detecting all samples. The precision and accuracy of the Y-chromosome target was lower comparing with the small and large autosomal targets. The threshold of 0.005ng/µl was established to proceed to DNA amplification. The internal validation conducted provided parameters for decisionmaking regarding the amount of human DNA required for amplification in the laboratory.
Keywords
DNA quantification, quality assurance, real time PCR, threshold, validation, Quantiplex® Pro