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VALIDATION OF AMPFLSTR® SGM PLUS® AMPLIFICATION KIT AT LOW DNA CONCENTRATIONS

Internal validation of new method is a mandatory step before its implementation into the everyday casework. Touch DNA analysis has become very popular, and scientists are nowadays struggling with very low DNA concentrations to generate a full DNA profile. The aim of this study was to validate AmpFlSTR® SGM Plus® PCR amplification kit at low DNA concentrations. Genomic DNA from buccal swab and epithelial cells deposited on clothes was isolated using Chelex. Genomic DNA content of each sample was determined by quantitative real-time polymerase chain reaction using Quantifiler® Human DNA Quantification Kit. AmpFlSTR® Control DNA 007, buccal swab and touch DNA were diluted as follows: 0.06 ng/mL, 0.04 ng/mL, 0.02 ng/mL and 0, 01 ng/mL. Each sample was amplified in pentaplicate using the AmpFlSTR® SGM Plus® PCR amplification kit. Statistical data analysis was performed using Excel. Results were presented as mean value of DNA concentrations and loci number. In all AmpFlSTR® Control DNA 007 dilutions, a full DNA profile was detected. In buccal swab samples, a full DNA profile was not detected at a concentration of 0.01 ng/mL. Moreover, in touch DNA samples a full DNA profile was detected at a concentration of 0.06 ng/mL. Partial DNA profiles were identified in other concentrations tested. The results of this study showed a lower sensitivity of validated PCR kit when amplifying low amounts of DNA. The more sensitive kits with a greater number of loci should be used in forensic casework due to their higher sensitivity and improved STR performance for forensic casework.

validation; AmpFlSTR® SGM Plus® PCR amplification kit; low DNA concentrations; forensic casework

Presentation number: FG13 Abstract number: ABS-201-ISABS-2013