Developmental validation of the PowerPlex® 18D System, a rapid STR multiplex for analysis of reference samples

Oostdik, Kathryn; French, Julie L.; Yet, Donald; Smalling, Briana B.; Nolde, Craig; Vallone, Peter M.; Butts, Erica L.R.; Hill, Carolyn R.; Kline, Margaret C.; Rinta, Theresa; Gerow, Amy M.; Allen, Stacey R.; Huber, Christopher K.; Teske, John; Krenke, Benjamin E.; Ensenberger, Martin G.; Fulmer, Patricia M.; Sprecher, Cynthia J.

doi:10.1016/j.fsigen.2012.07.008

Cited by 27 publications

(9 citation statements)

References 16 publications

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“…However, most of these studies evaluated fresh blood samples which are applied immediately on the FTA cards, although other sample types also have been reported [11,[15][16][17]. Many of these studies have targeted forensically-relevant STR markers [18,19] or single nucleotide polymorphisms (SNPs) [20], and only few studies report the results from genome wide association studies [21]. DNA quality and quantity in PM samples with variable PM intervals (PMIs) stored on FTA paper have not tested to determine if such samples meet the demands of forensic or other genomic DNA analyses.…”

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confidence: 99%

DNA quality and quantity from up to 16 years old post-mortem blood stored on FTA cards

Rahikainen

Palo

Leeuw

et al. 2016

Forensic Science International

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Blood samples preserved on FTA cards offer unique opportunities for genetic research. DNA recovered from these cards should be stable for long periods of time. However, it is not well established as how well the DNA stored on FTA card for substantial time periods meets the demands of forensic or genomic DNA analyses and especially so for from post-mortem (PM) samples in which the quality can vary upon initial collection. The aim of this study was to evaluate the time-dependent degradation on DNA quality and quantity extracted from up to 16 years old post-mortem bloodstained FTA cards. Four random FTA samples from eight time points spanning 1998 to 2013 (n=32) were collected and extracted in triplicate. The quantity and quality of the extracted DNA samples were determined with Quantifiler(®) Human Plus (HP) Quantification kit. Internal sample and sample-to-sample variation were evaluated by comparing recovered DNA yields. The DNA from the triplicate samplings were subsequently combined and normalized for further analysis. The practical effect of degradation on DNA quality was evaluated from normalized samples both with forensic and pharmacogenetic target markers. Our results suggest that (1) a PM change, e.g. blood clotting prior to sampling, affects the recovered DNA yield, creating both internal and sample-to-sample variation; (2) a negative correlation between the FTA card storage time and DNA quantity (r=-0.836 at the 0.01 level) was observed; (3) a positive correlation (r=0.738 at the level 0.01) was found between FTA card storage time and degradation levels. However, no inhibition was observed with the method used. The effect of degradation was manifested clearly with functional applications. Although complete STR-profiles were obtained for all samples, there was evidence of degradation manifested as decreased peak heights in the larger-sized amplicons. Lower amplification success was notable with the large 5.1 kb CYP2D6 gene fragment which strongly supports degradation of the stored samples. According to our results, DNA stored on FTA cards is rather stable over a long time period. DNA extracted from this storage medium can be used as human identification purposes as the method used is sufficiently sensitive and amplicon sizes tend to be <400 bp. However, DNA integrity was affected during storage. This effect should be taken into account depending on the intended application especially if high quality DNA and long PCR amplicons are required.

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confidence: 99%

DNA quality and quantity from up to 16 years old post-mortem blood stored on FTA cards

Rahikainen

Palo

Leeuw

et al. 2016

Forensic Science International

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“…In a typical forensic DNA laboratory, direct polymerase chain reaction (PCR) amplification is commonly employed to overcome these issues (Myers et al 2012;Tucker et al 2012). Direct amplification of STR loci reduces the time required for sample preparation and potentially helps laboratories to process increased number of samples (Wang et al 2011;Oostdik et al 2013;Hall and Roy 2014). In addition, the direct PCR amplification technique may also reduce risks of cross-over contamination (Caputo et al 2017;Ambers et al 2018).…”

Section: Introductionmentioning

confidence: 99%

Assessment of QIAGEN™ Investigator® 24plex GO! kit workflow for autosomal STR profiling of forensic reference samples

Hakim

Khan²,

Ismail³

et al. 2020

Egypt J Forensic Sci

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Background DNA profiling has proven to be a valuable technique for identification of individuals in crime. Currently, the technique targets several short tandem repeat (STR) regions in human genome. However, increasing number of samples submitted for STR analysis may lead to delays due to the limited number of experienced analysts who might be available at any given moment and the time taken to complete lengthy DNA profiling procedures. This study was conducted to test the specificity, repeatability, reproducibility and robustness of Investigator® 24plex GO! kit for genotyping of reference samples submitted to the Royal Malaysian Police Forensic DNA Laboratory for DNA database. Material and methods In this study, Investigator® 24plex GO! kit was used to directly amplify STR loci from buccal swab cell of reference samples that had previously been STR typed using GlobalFiler™ Express kit. Capillary electrophoresis was carried out on a 3500xL Genetic Analyser using POP-4® Polymer. Amplified products were assigned to particular STR alleles using the GeneMapper ID-X version 1.4 software. Results Our study shows that STR profiles generated using Investigator® 24plex GO! gave concordance results with those previously obtained using the GlobalFiler™ Express kit. In addition, quality sensors included in the kit are of particular importance for determining the effectiveness of the PCR reaction and help to indicate the nature and quantity of DNA template for PCR amplification. Conclusion The Investigator® 24plex GO! kit is reliable for STR typing of reference samples.

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“…The document highlights key rules to follow by Forensic DNA laboratories in order to evaluate reliability, sensitivity, specificity, stability, and reproducibility of their methods. Unlike mRNA profiling, many validation experiments have been carried out on DNA marker based multiplex assay (Ensenberger et al, 2010;Oostdik et al, 2013;Oostdik et al, 2014). The authors have reported specifically validation of DNA multiplex kits with addition of more loci to the 13 core CODIS loci, enhanced buffer system with hot-start Taq DNA polymerase, sensitivity, concordance, inhibition tolerance amongst other criteria, following SWGDAM developmental validation guidelines; all these have made DNA typing more efficient and less time consuming.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of genetic markers for forensic identification of human body fluids

Afolabi

Roeder

Iyengar

et al. 2017

Forensic Science International: Genetics Supplement Series

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Developmental validation of the PowerPlex® 18D System, a rapid STR multiplex for analysis of reference samples

Cited by 27 publications

References 16 publications

DNA quality and quantity from up to 16 years old post-mortem blood stored on FTA cards

DNA quality and quantity from up to 16 years old post-mortem blood stored on FTA cards

Assessment of QIAGEN™ Investigator® 24plex GO! kit workflow for autosomal STR profiling of forensic reference samples

Evaluation of genetic markers for forensic identification of human body fluids

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