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The importance of forensic storage support: DNA quality from 11-year-old saliva on FTA cards

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Abstract

Storage conditions influence the integrity of the recoverable DNA from forensic evidence in terms of yield and quality. FTA cards are widely used in the forensic practice as their chemically treated matrix provides protection from the moment of collection to the point of analysis with current STR typing technology. In this study, we assess the recoverability and the integrity of DNA from 11-year-old saliva on FTA cards using a forensic quantitative real-time polymerase chain reaction (qPCR) commercial assay. The quality after long-term storage was investigated in order to evaluate if the FTA device could assure enough stability over time, applying some internally validated quality criteria of the STR profile. Furthermore, we used a 3D interpolation model to combine the quantitative and qualitative data from qPCR to calculate the minimum optimal DNA input (MODI) to add to the downstream PCR reaction based on the quantitative and qualitative data of a sample. According to our results, when saliva sample is properly transferred onto FTA cards and then correctly stored according to the manufacturer’s instructions, it is possible to recover sufficient amounts of DNA for human identification even after more than a decade of storage at ambient temperature. Degradation affected the quality of results especially when the Degradation Index exceeds the value of 2.12, requiring modifications of the standard internal workflow to improve the genotyping quality. Above this value, the application of a “corrective factor” to the PCR normalization process was necessary in order to adjust the recommended manufacturer’s PCR DNA input taking into account the degradation level. Our results demonstrated the importance to consider in predictive terms the parameters obtained with the real-time quantification assay, both in terms of quantity (DNA concentration) and of quality (DI, inhibition). Informatics predictive tools including qPCR data together with the variables of storage duration and conditions should be developed in order to optimize the DNA analysis process.

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References

  1. Vernarecci S, Ottaviani E, Agostino A, Mei E, Calandro L, Montagna P (2015) Quantifiler Trio Kit and forensic samples management: a matter of degradation. Forensic Sci Int Genet 16:77–85

    Article  CAS  Google Scholar 

  2. Alaeddini R, Walsh SJ, Abbas A (2010) Forensic implications of genetic analyses from degraded DNA—a review. Forensic Sci Int Genet 4:148–157

    Article  CAS  Google Scholar 

  3. Davis DL, O’brien EP, Bentzley CM (2000) Analysis of the degradation of oligonucleotide strands during the freezing/thawing processes using MALDI-MS. Anal Chem 15:5092–5096

    Article  Google Scholar 

  4. Kampmann ML, Buchard A, Børsting C, Morling N (2016) High-throughput sequencing of forensic genetic samples using punches of FTA cards with buccal swabs. Biotechniques. 61:149–151

    Article  CAS  Google Scholar 

  5. Green H, Tillmar A, Pettersson G, Montelius K (2019) The use of FTA cards to acquire DNA profiles from postmortem cases. Int J Legal Med. https://doi.org/10.1007/s00414-019-02015-2

    Article  Google Scholar 

  6. Mundorff AZ, Amory S, Huel R, Bilić A, Scott AL, Parsons TJ (2018) An economical and efficient method for postmortem DNA sampling in mass fatalities. Forensic Sci Int Genet 36:167–175

    Article  CAS  Google Scholar 

  7. GE Healthcare life sciences, Application note 29-0082-33 AA, STR amplification of DNA from buccal and blood samples stored long-term on Whatman FTA cards (2011) Available from: https://www.gelifesciences.com/en/it/solutions/Forensics/Products%20and%20Technology/Sample%20Purification%20Transport%20and%20Storage/Long-term%20and%20room%20temperature%20storage

  8. Rahikainen AL, Jukka UP, de Leeuw W, Budowle B, Sajantila A (2016) DNA quality and quantity from up to 16 years old post-mortem blood stored on FTA cards. Forensic Sci Int 261:148–153

    Article  CAS  Google Scholar 

  9. Hedell R, Hedman J, Mostad P (2018) Determining the optimal forensic DNA analysis procedure following investigation of sample quality. Int J Legal Med 132:955–966

    Article  Google Scholar 

  10. Ewing MM, Thompson JM, McLaren RS, Purpero VM, Thomas KJ, Dobrowski PA, DeGroot GA, Romsos EL, Storts DR (2016) Human DNA quantification and sample quality assessment: developmental validation of the PowerQuant system. Forensic Sci Int Genet. 23:166–177

    Article  CAS  Google Scholar 

  11. PowerQuant System Technical Manual, 3/18. Available at: http://www.promega.com/resources/protocols/technical-manuals/101/powerquant-system-protocol/ (Accessed 14.03.19)

  12. AmpFlSTR Identifiler Plus PCR Amplification Kit User Guide Rev F/15. Available at: https://www.thermofisher.com/order/catalog/product/4427368 (accessed 14.03.19)

  13. Hansson O, Gill P, Egeland T (2014) STR-validator: an open source platform for validation and process control. Forensic Sci Int Genet 13:154–166

    Article  CAS  Google Scholar 

  14. Butler J (2012) Advanced topics in forensic DNA typing: methodologies. Academic Press, Massachusetts

    Google Scholar 

  15. Quality Assurance Standards for Forensic DNA Testing Laboratories. Available at: https://www.fbi.gov/file-repository/quality-assurance-standards-for-forensic-dna-testing-laboratories.pdf/view (Accessed 14.03.19)

  16. GE Healthcare Life Sciences, Application note 28-9822-22 AA, Reliable extraction of DNA from Whatman™ FTA™ cards (2010) Available from: https://us.vwr.com/assetsvc/asset/en_US/id/16147319/contents

  17. Applied Biosystems, AmpFlSTR® Identifiler® Plus PCR Amplification Kit User Guide, Revision F

  18. Balding DJ (2013) Evaluation of mixed-source, low-template DNA profiles in forensic science. Proc Natl Acad Sci U S A 110:12241–12246

    Article  CAS  Google Scholar 

  19. Huang LH, Lin PH, Tsai KW, Wang LJ, Huang YH, Kuo HC, Li SC (2017) The effects of storage temperature and duration of blood samples on DNA and RNA qualities. PLoS One 12(9):e0184692. https://doi.org/10.1371/journal.pone.0184692

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Badu-Boateng A, Twumasi P, Salifu SP, Afrifah KA (2018) A comparative study of different laboratory storage conditions for enhanced DNA analysis of crime scene soil-blood mixed sample. Forensic Sci Int 292:97–109

    Article  CAS  Google Scholar 

  21. Lin S, Li C, Ip SCY (2018) A performance study on three qPCR quantification kits and their compatibilities with the 6-dye DNA profiling systems. Forensic Sci Int Genet. 33:72–83

    Article  Google Scholar 

  22. Hansson O, Egeland T, Gill P (2017) Characterization of degradation and heterozygote balance by simulation of the forensic DNA analysis process. Int J Legal Med 131:303–317

    Article  Google Scholar 

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Authors and Affiliations

  1. Department of Biomedical, Metabolic and Neural Sciences, Institute of Legal Medicine, University of Modena and Reggio Emilia, Modena, Italy

    Beatrice Corradini, Milena Alù & Enrico Silingardi

  2. Department of Mathematics, University of Bologna, Bologna, Italy

    Elena Magnanini

  3. Department of Physics, Informatics and Mathematics, University of Modena and Reggio Emilia, Modena, Italy

    Mathilde Emmanuelle Galinier

Authors
  1. Beatrice Corradini
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  2. Milena Alù
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  3. Elena Magnanini
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  4. Mathilde Emmanuelle Galinier
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  5. Enrico Silingardi
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Corresponding author

Correspondence to Beatrice Corradini.

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Corradini, B., Alù, M., Magnanini, E. et al. The importance of forensic storage support: DNA quality from 11-year-old saliva on FTA cards. Int J Legal Med 133, 1743–1750 (2019). https://doi.org/10.1007/s00414-019-02146-6

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  • DOI: https://doi.org/10.1007/s00414-019-02146-6

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