Abstract
The Investigator® Quantiplex HYres kit was evaluated as a potential replacement for dual DNA quantification
of casework samples. This kit was determined to be highly sensitive with a limit of
quantification and limit of detection of 0.0049 ng/μL and 0.0003 ng/μL, respectively, for both human and male DNA, using full or half reaction volumes.
It was also accurate in assessing the amount of male DNA present in 96 mock and actual
casework male:female mixtures (various ratios) processed in this exercise. The close
correlation between the male/human DNA ratios expressed in percentages derived from
the Investigator® Quantiplex HYres quantification results and the male DNA proportion calculated in
mixed AmpFlSTR® Profiler® Plus or AmpFlSTR® Identifiler® Plus profiles, using the Amelogenin Y peak and STR loci, allowed guidelines to be
developed to facilitate decisions regarding when to submit samples to Y-STR rather
than autosomal STR profiling. The internal control (IC) target was shown to be more
sensitive to inhibitors compared to the human and male DNA targets included in the
Investigator® Quantiplex HYres kit serving as a good quality assessor of DNA extracts. The new
kit met our criteria of enhanced sensitivity, accuracy, consistency, reliability and
robustness for casework DNA quantification.
Keywords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Forensic Science International: GeneticsAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- A simple and sensitive method for quantifying human genomic DNA in forensic specimen extracts.BioTechniques. 1989; 7: 852-855
- Sensitive and specific quantification of human genomic deoxyribonucleic acid (DNA) in forensic science specimens: casework examples.J. Forensic Sci. 1991; 36: 1198-1203
- NIST standards development for RFLP DNA profiling.in: Proceedings from Promega 2nd Int. Symp. Hum. Ident., Promega Corporation, Scottsdale, AZ, October 12–151991: 245-261
- A rapid chemiluminescent method for quantitation of human DNA.Nucleic Acids Res. 1992; 20: 5061-5065
- Characterization of human AFLP systems apolipoprotein B, phenylalanine hydroxylase, and D1S80.PCR Methods Appl. 1994; 3: 351-358
- Development, characterization, and validation of a sensitive primate-specific quantification assay for forensic analysis.BioTechniques. 2003; 34: 314-322
- Real-time DNA quantification of nuclear and mitochondrial DNA in forensic analysis.BioTechniques. 2002; 33: 402-411
- Developmental validation of a real-time quantitative PCR assay for automated quantification of human DNA.J. Forensic Sci. 2003; 48: 1041-1046
- Rapid quantification and sex determination of forensic evidence materials.J. Forensic Sci. 2003; 48: 1280-1287
- Real-time PCR designs to estimate nuclear and mitochondrial DNA copy number in forensic and ancient DNA studies.Forensic Sci. Int. 2004; 139: 141-149
B.E. Krenke, S., Ekenberg, S., Frackman, K., Hoffmann, C.J., Sprecher, D.R. Storts, Development of a novel, fluorescent, two-primer approach to quantitative PCR, Profiles in DNA, April 2005, pp. 3–5.
- Developmental validation of the Quantifiler™ real-time PCR kits for the quantification of human nuclear DNA samples.J. Forensic Sci. 2005; 50: 809-825
- A duplex real-time qPCR assay for the quantification of human nuclear and mitochondrial DNA in forensic samples: implications for quantifying DNA in degraded samples.J. Forensic Sci. 2005; 50: 1044-1060
- Multiplex polymerase chain reaction for simultaneous quantitation of human nuclear, mitochondrial, and male Y-chromosome DNA: application in human identification.Anal. Biochem. 2005; 337: 89-97
- A quantitative PCR assay for the assessment of DNA degradation in forensic samples.Forensic Sci. Int. 2006; 158: 14-26
- A quadruplex real-time qPCR assay for the simultaneous assessment of total human DNA, human male DNA, DNA degradation and the presence of PCR inhibitors in forensic samples: A diagnostic tool for STR typing.Forensic Sci. Int. Genet. 2008; 2: 108-125
- Development of a real-time method to detect DNA degradation in forensic samples.J. Forensic Sci. 2012; 57: 466-471
- Developmental validation of a real-time PCR assay for the simultaneous quantification of total human and male DNA.Forensic Sci. Int. Genet. 2008; 3: 14-21
- Developmental validation of the Quantifiler® Duo DNA Quantification Kit for simultaneous quantification of total human and human male DNA and detection of PCR inhibitors in biological samples.J. Forensic Sci. 2009; 54: 305-319
- Quantifiler® Duo DNA Quantification Kit: a guiding tool for short tandem repeat genotyping of forensic samples.J. Forensic Res. 2011; 2: 118https://doi.org/10.4172/2157-7145.1000118
- Selective quantification of human DNA by real-time PCR of FOXP2.Forensic Sci. Int. Genet. 2012; 6: 447-451
- Quantitative real-time PCR (qPCR) assay for human-dog-cat species identification and nuclear DNA quantification.Forensic Sci. Int. Genet. 2012; 6: 290-295
- Quadriplex real-time PCR (qPCR) assay for human-canine-feline species identification and nuclear DNA quantification.Forensic Sci. Int. Genet. 2012; 6: e97-e98
- Improving STR analysis using a novel quantification technology—more than simply DNA quantification.in: Poster #35 from 6th Eur. Acad. Forensic Sci. Conf., The Hague, The Netherlands, August 20–242012
- Qiagen’s Investigator™ Quantiplex Kit as a predictor of STR amplification success from low-yield DNA samples.J. Forensic Sci. 2013; 58: 1306-1309
- Prime time for real-time PCR.Nat. Methods. 2005; 2: 305-312
- The use of real-time PCR methods in DNA sequence variation analysis.Clin. Chim. Acta. 2006; 363: 32-47
- Detection of PCR products using self-probing amplicons and fluorescence.Nat. Biotechnol. 1999; 17: 804-807
- Design and use of scorpions fluorescent signaling molecules, Methods Mol. Biol. 429 99–115.in: Marx A. Steiz O. Molecular Beacons: Signalling Nucleic Acid Probes, Methods and Protocols. Humana Press Inc., Totowa, NJ2008
- Multiplex real-time PCR assay using Scorpion probes and DNA capture for genotype-specific detection of Giardia lamblia on fecal samples.J. Clin. Microbiol. 2005; 43: 1256-1260
- Real-time PCR detection and speciation of Cryptosporidium infection using scorpion probes.J. Med. Microbiol. 2006; 55: 1217-1222
- Development of a novel quantitative real-time assay using duplex Scorpion primer for detection of Chlamydia trachomatis.Exp. Mol. Pathol. 2007; 83: 119-124
- Quadruplex real-time PCR assay using allele-specific Scorpion primers for detection of mutations conferring clarithromycin resistance to Helicobacter pylori.J. Clin. Microbiol. 2008; 46: 2320-2326
- Development of a scorpion probe-based real-time PCR for the sensitive quantification of Bacteroides sp. ribosomal DNA from human and cattle origin and evaluation in spring water matrices.Microbiol. Res. 2008; 163: 140-147
- Real-time PCR detection of Mycoplasma pneumoniae in the diagnosis of community-acquired pneumonia, Methods Mol. Biol. 1160 99–105.in: Biassoni R. Raso A. Quantitative Real-Time PCR: Methods and Protocols. Springer Science + Business Media, New York, NY2014
- Quantitation of HIV-1 by real-time PCR with a unique fluorogenic probe.J. Virol. Methods. 2001; 93: 33-42
- Real-time quantitative assay of HCV RNA using the duplex scorpion primer.Arch. Virol. 2007; 152: 431-440
- Real-time quantitative PCR assays for quantification of L1781 ACCase inhibitor resistance allele in leaf and seed pools of Lolium populations.Pest Manage. Sci. 2006; 62: 1082-1091
- Mode of action and application of scorpion primers to mutation detection.Nucleic Acids Res. 2000; 28: 3752-3761
- Multiplexed assays for detection of mutations in PIK3CA.Clin. Chem. 2008; 54: 757-760
- Development of a human DNA quantitation system.Croat. Med. J. 2001; 42: 336-339
- A more sensitive method for the quantitation of genomic DNA by Alu amplification.J. Forensic Sci. 2002; 47: 589-592
- Development of an Alu-based, real-time PCR method for quantitation of human DNA in forensic samples.J. Forensic Sci. 2003; 48: 936-944
- Simultaneous determination of total human and male DNA using a duplex real-time PCR assay.J. Forensic Sci. 2006; 51: 1005-1015
- Human genomic DNA quantitation system, H-Quant: development and validation for use in forensic casework.J. Forensic Sci. 2007; 52: 364-370
- Inter-laboratory testing of a highly sensitive quantification system for assessing DNA quality in forensic samples.in: Poster A53 from 66th Amer. Acad. Forensic Sci. Conf., Seattle, WA, February 17–222014
- Development of a highly sensitive human and male quantification system for assessing DNA quality in forensic samples.in: Poster A67 from 66th Amer. Acad. Forensic Sci. Conf., Seattle, WA, February 17–222014
Investigator® Quantiplex HYres Handbook. Version dated March 2012. QIAGEN, Hilden, Germany.
QIAGEN Validation Report. Developmental validation of the Investigator® Quantiplex HYres Kit. Version dated 11/2012.
- An accelerated analytical process for the development of STR profiles for casework samples.J. Forensic Sci. 2015; (in press)
- Optimized configuration of fixed-tip robotic liquid-handling stations for the elimination of biological sample cross-contamination.JALA. 2007; 12: 339-354
- Automated processing of forensic casework samples using robotic workstations equipped with nondisposable tips: contamination prevention.J. Forensic Sci. 2008; 53: 632-651
- Validation of a DNA IQ™-based extraction method for TECAN robotic liquid handling workstations for processing casework.Forensic Sci. Int. Genet. 2010; 4: 292-304
- Developmental and usage of a NIST standard reference material for real time PCR quantitation of human DNA.in: Poster #303 from 22nd Int. Soc. Forensic Genet. Congress, Copenhagen, Denmark, September 21–252007
- Developmental and usage of a NIST standard reference material for real time PCR quantitation of human DNA.Forensic Sci. Int. Genet. Suppl. Ser. 2008; 1: 80-82
- Using SRM 2372 Human DNA Quantitation Standard, Are there differences between qPCR assays.in: Poster #49 from Promega 20th Int. Symp. Hum. Ident., Promega Corporation, Las Vegas, NV, October 12–152009
- Production and certification of NIST Standard Reference Material 2372 Human DNA Quantitation Standard.Anal. Bioanal. Chem. 2009; 394: 1183-1192
- Re-certification of NIST Standard Reference Material® 2372 Human DNA Quantitation Standard: the what, the why and the how.in: Proceedings A174 from 65th Amer. Acad. Forensic Sci. Ann. Scientific Meeting, Washington, DC, February 18–232013
Quantifiler® Duo DNA Quantification Kit User’s Manual, Applied Biosystems, ©2008, 01/2008 Rev. A pp.1–6.
Article info
Publication history
Published online: December 30, 2014
Accepted:
December 27,
2014
Received:
November 12,
2014
Identification
Copyright
© 2015 Published by Elsevier Inc. All rights reserved.
