Rapid PCR of STR markers: Applications to human identification

Romsos, Erica L.; Vallone, Peter M.

doi:10.1016/j.fsigen.2015.04.008

Cited by 49 publications

(35 citation statements)

References 110 publications

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“…Integration of these steps into a single instrument is a challenging goal but has become possible in recent years. Over the last decade, there have been many advances to reduce time within the DNA typing process and the laboratory‐sized footprint . These advances include rapid PCR protocols for short tandem repeat (STR) typing , advancements in microfluidic efforts for PCR , and separation and detection .…”

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confidence: 99%

Results of the 2018 Rapid DNA Maturity Assessment

Romsos

French²,

Smith³

et al. 2020

Journal of Forensic Sciences

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Three commercially available integrated rapid DNA instruments were tested as a part of a rapid DNA maturity assessment in July of 2018. The assessment was conducted with sets of blinded single-source reference samples provided to participants for testing on the individual rapid platforms within their laboratories. The data were returned to the National Institute of Standards and Technology (NIST) for review and analysis. Both FBI-defined automated review (Rapid DNA Analysis) and manual review (Modified Rapid DNA Analysis) of the datasets were conducted to assess the success of genotyping the 20 Combined DNA Index System (CODIS) core STR loci and full profiles generated by the instruments. Genotype results from the multiple platforms, participating laboratories, and STR typing chemistries were combined into a single analysis. The Rapid DNA Analysis resulted in a success rate of 80% for full profiles (85% for the 20 CODIS core loci) with automated analysis. Modified Rapid DNA Analysis resulted in a success rate of 90% for both the CODIS 20 core loci and full profiles (all attempted loci per chemistry). An analysis of the peak height ratios demonstrated that 95% of all heterozygous alleles were above 59% heterozygote balance. For base-pair sizing precision, the precision was below the standard 0.5 bp deviation for both the ANDE 6C System and the RapidHIT 200.

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confidence: 99%

Results of the 2018 Rapid DNA Maturity Assessment

Romsos

French²,

Smith³

et al. 2020

Journal of Forensic Sciences

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“…This is not including subsequent data analysis, which can take as little as 5 minutes, using GeneMapper IDX as an expert system with our internally validated settings. This process is of a similar timescale to other published studies that have tested standard DNA profiling pipelines with FTA cards for reference profiling [5]. All samples processed through this method produced full DNA profiles in comparison to reference data within a 97-102 minute timescale, and these results were used as a baseline comparison to the results produced with altered settings by comparing profile quality and average peak height across all alleles.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to improvements made by STR kit manufacturers, published studies looking at reducing the time taken to generate a DNA profile using standard laboratory equipment have focused on use of direct PCR for touch and trace DNA [5,6], use of enhanced DNA polymerase enzymes [5,7,8,9,10], modified PCR mixes [9,11], and use of more efficient thermal cycler instrumentation [10,12]. In addition, research has also been carried out in the field of microfluidic chip systems to separate and detect fragments in place of traditional capillary electrophoresis platforms [5,13].…”

Section: Introductionmentioning

confidence: 99%

Optimisation of rapid STR analysis using a standard DNA forensic pipeline

Gammon

Mayers

2019

Preprint

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Previous studies in published literature have reported on various alterations to STR mastermixes, protocols and instrumentation in order to reduce the time taken to generate forensic DNA profiles from reference and casework type samples. In this study, we demonstrate how altering default PCR amplification and capillary electrophoresis protocols in our existing DNA profiling pipeline can reduce the overall time taken to generate a DNA profile from buccal cell reference samples. GlobalFiler Express STR mastermix was used with direct PCR from FTA cards, run on altered PCR protocols and CE settings, and results compared to the standard evaluated settings used in our laboratories. This study demonstrated that full DNA profiles could be recovered in less than 80 minutes in comparison to our standard time of 97 -102 minutes whilst utilising existing reagent kits and instrumentation, with only minor modifications to protocols.Rapid DNA | direct amplification | forensic DNA analysis | STR profiling

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“…The uses of fluorophores sensitized by fluorescence resonance energy transfer (FRET) provided detection kinetics of fluorescence accumulation, and combined PCR amplification with real-time detection [76,77,99,100]. Alternative, or engineered, high-processivity DNA polymerases can have increased resistance to inhibitory factors relating to amplification of target DNA directly from samples without prior extraction or quantification [73,78,101,102].…”

Section: Extraction Of Compound And/or Challenging Samplesmentioning

confidence: 99%

Forensic DNA Technological Advancements as an Emerging Perspective on Medico-Legal Autopsy: A Mini Review

Pádár¹,

Zenke²,

Kozma³

2018

Post Mortem Examination and Autopsy - Current Issues From Death to Laboratory Analysis

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The importance of biological traces and evidences related to a criminal matter has been recognized for a long time. The examination of the expression of genetic polymorphism has been an integral part of the multidisciplinary field of medico-legal autopsy for over a century. Since the initial application of blood group antigens for personalization of a putative perpetrator in a murder case, the discipline of forensic genetics has evolved as a standard of forensic sciences. The real breakthrough, the application of molecular tools and processes for the in-vitro replication of genetic substances, has increasingly allowed the exploitation of advances of molecular genetics for both forensic and criminal investigations. Although there are certainly many more applications and scientific fields in the medico-legal arena, the relatively fast progress of genetics, which has accelerated recently with state-of-art technologies, can provide ever more relevant information in relation to a corpse or the cause and manner that resulted in the corpse for autopsy. This topic concerns the currently accepted forensic DNA technology, and the last section reviews commonly used markers for nuclear and mitochondrial DNA analysis as well as ongoing research. This review also focuses on the increasingly important non-human sources of DNA, and shortly covers the main aspects of animal forensic DNA examination.

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Rapid PCR of STR markers: Applications to human identification

Cited by 49 publications

References 110 publications

Results of the 2018 Rapid DNA Maturity Assessment

Results of the 2018 Rapid DNA Maturity Assessment

Optimisation of rapid STR analysis using a standard DNA forensic pipeline

Forensic DNA Technological Advancements as an Emerging Perspective on Medico-Legal Autopsy: A Mini Review

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