Operationalisation of the ForenSeq® Kintelligence Kit for Australian unidentified and missing persons casework.

Single nucleotide polymorphism (SNP) genotyping technologies can generate investigative leads for human remains identification, including estimation of biological sex, biogeographical ancestry (BGA), externally visible characteristics (EVCs), identity, uniparental lineage and extended kinship. The ForenSeq® Kintelligence Kit provides forensic laboratories with the ability to apply this suite of genetic tools to forensic samples using one panel targeting 10,230 SNPs (including 56 ancestry-informative, 24 phenotype-informative, 94 identity-informative, 106 X chromosome, 85 Y chromosome and 9867 kinship-informative SNPs) sequenced on the MiSeq FGx® Sequencing System. The ForenSeq® Kintelligence Kit has been internally validated, optimised and operationalised by the Australian Federal Police National DNA Program for Unidentified and Missing Persons (AFP Program) for coronial casework. The internal validation was conducted according to the Scientific Working Group on DNA Analysis Methods guidelines (excluding mixture analysis), focussing on sample types typically encountered in human remains identification casework, such as bones, teeth, nail, blood and hair. The workflow was optimised for a high throughput library preparation and sequencing workflow, and additional analytical thresholds were developed to improve genotyping accuracy for low DNA input samples. Additionally, the genetic intelligence generated from the kit was compared to the self-declared biological sex, EVCs and BGA of the DNA donors to assess concordance. The kit was able to produce high quality SNP profiles from 1.0 ng down to 0.1 ng of DNA, with high repeatability and reproducibility, and minimal background noise. The prediction accuracy for biological sex (95%), hair colour (58%), eye colour (74%) and BGA inferences (consistent: 74%; partially consistent: 10%; inconclusive: 16%) was determined based on self-declared data. Additionally, SNP profiles from a volunteer family group of ten related individuals were uploaded to GEDmatch PRO™ to assess kinship accuracy. The kit was capable of detecting (97%) and accurately classifying (90%) genetic relationships spanning from first to fifth degree. The Kintelligence Kit provides the AFP Program with a robust and reliable genetic intelligence tool for unidentified and missing persons investigations, which has been designed to sequence multiple challenging samples in a single multiplexed assay using existing laboratory instrumentation.

Corrigendum: Dreading Yet Hoping: Traumatic Loss Impacted by Reference DNA Sample Collection for Families of Missing People.

[This corrects the article DOI: 10.3389/fpsyt.2022.866269.].

Dreading Yet Hoping: Traumatic Loss Impacted by Reference DNA Sample Collection for Families of Missing People.

The trauma of having a family member missing is commonly described as an ambiguous loss where the finality of the loss is not realized, as is experienced with a death. There is uncertainty due to the trauma of the absence and subsequent police investigation, leading to physical and emotional impacts for the aftercare of those left behind. There are 850 unidentified human remains and 2,600 long-term missing persons cases in Australia. The Australian Federal Police (AFP) National DNA Program for Unidentified and Missing Persons aims to scientifically link these cases using modern DNA techniques and databases. A DNA-led identification effort may assist to provide answers to Australian families searching for missing relatives, but may also contribute to the trauma experienced by these families. A literature review demonstrated empirical research for the development of scientific best practices for the collection of reference DNA samples for forensic purposes, but minimal evidence about the impact of reference DNA sample collection on kin when attempting to identify the deceased remains of missing people in non-mass casualty situations. The aim of this study was to develop an academically robust understanding of the unique impact of reference DNA sample collection on families of missing persons and support pathways tailored to the experience. This study involved 26 Australian families of long-term missing (ranging from 1 to 20+ years) people in Australia anonymously completing a mixed-methods online survey about their experiences of providing reference DNA samples to aid missing persons investigations. Respondents were representative of a range of ages, genders and relationships to the missing individual. The thematic analysis of the survey results identified the provision of a reference DNA sample: (1) resembles an overt act of hope as families perceive their sample assists the investigation, whilst also being traumatic, triggered by the prospect of scientifically matching their missing family member to a set of unknown human remains; (2) can cause immediate interpersonal impacts and ongoing impacts to families' wellbeing; and (3) can be improved by considering the environment where the sample is collected, professionalism of the police officer collecting the sample, timeliness of the provision of the sample, level of support provided during and after sample collection, and effective communication of forensic procedures and processes as they relate to the missing persons investigation. The study concludes that the complexity associated with provision of family reference samples requires the development and implementation of best practice guidelines, including psycho-education strategies to be used by practitioners to minimize the vicarious trauma for relatives already traumatized by the loss of their missing family member. These guidelines would support the objectives of the AFP Program and benefit all routine missing persons investigations.

Psychological distress, depression, anxiety, stress, and exercise in Australian and New Zealand mothers: A cross-sectional survey.

Postnatal psychological well-being is a complex issue characterized by major changes in physical, social, and emotional health. In response to addressing limitations of previous research this study aimed to: (i) quantify psychological distress, depression, anxiety, and stress in a large sample of Australian and New Zealand mothers and to compare to Australian norm data; and (ii) identify any significant differences in psychological distress, depression, anxiety, and stress of the mothers when grouped according to their self-reported exercise incidence. Self-reported short-form version Depression Anxiety Stress Scale data were collected from 3601 Australia and New Zealand mothers with at least one child under the age of 5 years. Results demonstrated that this sample of mothers had significantly poorer psychological well-being than the general Australian population. Furthermore, with the exception of anxiety, psychological well-being of mothers who reported exercising three to four times per week was significantly and meaningfully more positive compared to those mothers that reported not to exercise.

A grass molecular identification system for forensic botany: a critical evaluation of the strengths and limitations.

Plant material is frequently encountered in criminal investigations but often overlooked as potential evidence. We designed a DNA-based molecular identification system for 100 Australian grasses that consisted of a series of polymerase chain reaction assays that enabled the progressive identification of grasses to different taxonomic levels. The identification system was based on DNA sequence variation at four chloroplast and two mitochondrial loci. Seventeen informative indels and 68 single-nucleotide polymorphisms were utilized as molecular markers for subfamily to species-level identification. To identify an unknown sample to subfamily level required a minimum of four markers or nine markers for species identification. The accuracy of the system was confirmed by blind tests. We have demonstrated "proof of concept" of a molecular identification system for trace botanical samples. Our evaluation suggests that the adoption of a system that combines this approach with DNA sequencing could assist the morphological identification of grasses found as forensic evidence.