Improved sample collection and preservation of DNA from decomposing human remains; A direct PCR approach for faster disaster victim identification (DVI)

As submitted by the proposer: In the event of a mass disaster, forensic scientists may be faced with the task of processing hundreds or even thousands of decomposing bodies for DNA identification. The enormity of the event often results in delayed recovery and processing of bodies. In remote and tropical locations, the delayed disaster victim identification (DVI) response and lack of resources such as refrigerated mortuaries quickly result in decomposition of the human remains. This leads to severe degradation of DNA in these tissues, making DNA typing more difficult. With advanced decomposition, skin and muscle tissue often remain, and are easily and quickly sampled. This project will use decomposing human cadavers (N=3) to demonstrate that skin and muscle samples can be collected quickly in the field, stored and adequately preserved for months in hostile climates (without refrigeration) in formats amenable to much faster processing for successful DNA typing.

This study will be divided into two phases and conducted over 36 months; 1) Optimization of TENT buffer to maximize the amount of "free" DNA in solution for direct amplification, and 2) Develop strategies for simple field collection and storage of decomposing tissues using FT A-based devices, coupled with fast direct amplification protocols.

Methods will be optimized based on matrices such as the amount of DNA (via qPCR) and quality of DNA recovered (Degradation Index in QuantiFiler® Trio, Quality Sensor and no. alleles successfully amplified using the Investigator 24plex QS STR kit) from samples stored for up to three months in harsh conditions. Data will be tested for statistical significance by ANOVA with Neumann-Keuls post-hoc comparisons, or Student t-test when appropriate. P<0.05 will be accepted as the level of significance.

The novel combination of sample collection methods in the field, tissue preservation and more rapid sample processing has tremendous potential for improving DNA profiling success and decrease time/cost for forensic analyses as compared with current DVI practices for DNA identification. Expected scholarly products will include a final technical report, publications in high-Impact factor journals and presentation of data at scientific meetings. In an effort to facilitate technology transition into practice, the most successful protocols developed in this study may be reported to the OSAC DVI Sub-committee as recommendations for improvement in tissue preservation and DNA processing methods which markedly increase throughput of human tissue samples in cases of mass fatality events.

This project contains a research and/or development component, as defined in applicable law. nca/ncf.

As submitted by the proposer: In the event of a mass disaster, forensic scientists may be faced with the task of processing hundreds or even thousands of decomposing bodies for DNA identification. The enormity of the event often results in delayed recovery and processing of bodies.

In remote and tropical locations, the delayed disaster victim identification (DVI) response and lack of resources such as refrigerated mortuaries quickly result in decomposition of the human remains. This leads to severe degradation of DNA in these tissues, making DNA typing more difficult. With advanced decomposition, skin and muscle tissue often remain, and are easily and quickly sampled. This project will use decomposing human cadavers (N=3) to demonstrate that skin and muscle samples can be collected quickly in the field, stored and adequately preserved for months in hostile climates (without refrigeration) in formats amenable to much faster processing for successful DNA typing.

This study will be divided into two phases and conducted over 36 months; 1) Optimization of TENT buffer to maximize the amount of "free" DNA in solution for direct amplification, and 2) Develop strategies for simple field collection and storage of decomposing tissues using FT A-based devices, coupled with fast direct amplification protocols.

Methods will be optimized based on matrices such as the amount of DNA (via qPCR) and quality of DNA recovered (Degradation Index in QuantiFiler® Trio, Quality Sensor and no. alleles successfully amplified using the Investigator 24plex QS STR kit) from samples stored for up to three months in harsh conditions. Data will be tested for statistical significance by ANOVA with Neumann-Keuls post-hoc comparisons, or Student t-test when appropriate. P<0.05 will be accepted as the level of significance.

The novel combination of sample collection methods in the field, tissue preservation and more rapid sample processing has tremendous potential for improving DNA profiling success and decrease time/cost for forensic analyses as compared with current DVI practices for DNA identification. Expected scholarly products will include a final technical report, publications in high-Impact factor journals and presentation of data at scientific meetings.

In an effort to facilitate technology transition into practice, the most successful protocols developed in this study may be reported to the OSAC DVI Sub-committee as recommendations for improvement in tissue preservation and DNA processing methods which markedly increase throughput of human tissue samples in cases of mass fatality events.

This project contains a research and/or development component, as defined in the applicable law. nca/ncf