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A performance study on three qPCR quantification kits and their compatibilities with the 6-dye DNA profiling systems
Sze-Wah Lin1, Christina Li1, Stephen C Y Ip1
1Forensic Science Division, Government Laboratory, Homantin Government Offices, Hong Kong, China.
Forensic Science International. Genetics
|December 5, 2017
Summary
New qPCR kits for forensic DNA quantification show improved precision and inhibition tolerance. Quantifiler® Trio and PowerQuant® systems offer better autosomal and male DNA quantification, with degradation index features for challenging samples.
Area of Science:
- Forensic Science
- Molecular Biology
- Genetics
Background:
- DNA quantification is crucial for forensic DNA profiling, estimating DNA amounts for optimal amplification and assessing extract quality.
- Advanced quantification systems can also estimate DNA degradation levels.
Purpose of the Study:
- To evaluate the performance and compatibility of three new generation quantitative PCR (qPCR) kits: QIAGEN's Investigator® Quantiplex HYres, Applied Biosystems' Quantifiler® Trio, and Promega's PowerQuant® System.
- To assess their performance with three 6-dye DNA profiling systems.
Main Methods:
- Performance evaluation of Investigator® Quantiplex HYres, Quantifiler® Trio, and PowerQuant® qPCR kits.
- Assessment of standard curve consistency, reproducibility, male DNA detection in excess female DNA, and PCR inhibition tolerance.
- Compatibility testing with GlobalFiler, Fusion 6C, and Investigator 24plex DNA profiling systems.
Main Results:
- All three kits demonstrated satisfactory consistency and reproducibility, effectively screening male DNA in excess female DNA.
- The kits showed improved tolerance to PCR inhibition compared to the Quantifiler® Human DNA Quantification Kit.
- PowerQuant® exhibited superior precision in both autosomal and male DNA quantification.
- Quantifiler® Trio and PowerQuant® showed better correlation between autosomal and male DNA quantification and include degradation index features.
- Quantifiler® Trio and PowerQuant® demonstrated good compatibility with GlobalFiler and Fusion 6C profiling systems, while Quantiplex HYres provided a reliable IPC for Investigator 24plex.
Conclusions:
- The evaluated new generation qPCR kits offer enhanced performance for forensic DNA quantification.
- Quantifiler® Trio and PowerQuant® provide more precise quantification and better compatibility with specific DNA profiling systems, including features for degraded or inhibited samples.
- Quantiplex HYres is suitable for inhibition detection with the Investigator 24plex system.
