Systematic optimisation of crude buccal swab lysate protocols for use with the ForenSeq™ DNA Signature Prep Kit.

The ForenSeq™ DNA Signature Prep kit has not been thoroughly tested with crude buccal swab lysates in large-scale population studies using massively parallel sequencing (MPS). Commonly used lysis buffers for swabs intending to undergo direct polymerase chain reaction (PCR) are SwabSolution™ and STR GO! Lysis Buffers, and these have been successfully used to generate population data using capillary electrophoresis (CE) systems. In this study, we investigated the performance and optimisation of SwabSolution™ and STR GO! lysates with the ForenSeq™ DNA Signature Prep workflow and addressed the challenge of failed MPS profiles in initial trials.

Disclosure of biological sex may impact individual privacy.

Human identification by forensic DNA profiling primarily relies on the analysis of short tandem repeat markers (STRs) and Amelogenin or other sex determining markers. The resultant DNA profiles can be compared directly between evidence and reference samples or indirectly (i.e.

Longitudinal assessment of DNA recovery from post-mortem whole blood stored in EDTA, sodium fluoride/potassium oxalate and additive-free tubes.

Adverse drug reactions and fatalities can result from therapeutic drug use due to genetic deficiencies in drug-metabolizing enzymes. In cases where ancillary testing may not reveal a clear cause of death, molecular autopsies can be valuable. However, forensic mortuaries do not retain DNA samples in all cases, which hinders subsequent genetic testing if it is later deemed necessary.

A 10-year retrospective analysis of sudden unexpected death in the young investigated at Salt River Mortuary, Cape Town.

Sudden unexpected death in the young (SUDY) is defined as the rapid, unsuspected demise of an apparently healthy individual between the ages of one and 40 years. There is a gap in research pertaining to this population in a South African context. This retrospective study aimed to explore the burden, scope of post-mortem investigation, and risk factors of SUDY admissions to Salt River Mortuary (SRM) in Cape Town between 1 January 2010 and 31 December 2019.

A study protocol to characterise pathophysiological and molecular markers of rheumatic heart disease and degenerative aortic stenosis using multiparametric cardiovascular imaging and multiomics techniques.

Introduction: Rheumatic heart disease (RHD), degenerative aortic stenosis (AS), and congenital valve diseases are prevalent in sub-Saharan Africa. Many knowledge gaps remain in understanding disease mechanisms, stratifying phenotypes, and prognostication. Therefore, we aimed to characterise patients through clinical profiling, imaging, histology, and molecular biomarkers to improve our understanding of the pathophysiology, diagnosis, and prognosis of RHD and AS.

Clinical exome sequencing elucidates underlying cause of death in sudden unexpected death of infants: two case reports.

Sudden unexpected death in infants (SUDI) is a traumatic event for families, and unfortunately its occurrence remains high in many parts of the world. Whilst cause of death is resolved for most cases, others remain undetermined following postmortem investigations. There has been a recognition of the role of genetic testing in unexplained cases, where previous studies have demonstrated the resolution of cases through DNA analyses.

Understanding the burden of unidentified bodies: a systematic review.

While human identification is a crucial aspect of medico-legal investigations, many individuals remain unidentified each year across the world. The burden of unidentified bodies is often referred to when motivating for improved methods of identification, and anatomical teaching, yet the actual burden is somewhat unclear. A systematic literature review was undertaken to identify articles that empirically investigate the number of unidentified bodies experienced.

Forensic DNA extraction methods for human hard tissue: A systematic literature review and meta-analysis of technologies and sample type.

DNA identification of human remains has a valuable role in the field of forensic science and wider. Although DNA is vital in identification of unknown human remains, post-mortem environmental factors can lead to poor molecular preservation. In this respect, focus has been placed on DNA extraction methodologies for hard tissue samples, as these are the longest surviving.

Forensic human identification: retrospective investigation of anthropological assessments in the Western Cape, South Africa.

The identification of unknown persons, particularly those who are decomposed or burnt, is a global challenge. Forensic Anthropology Cape Town (FACT) is a service provider that assists the South African state with the identification of human remains. However, empirical data pertaining to anthropologically analysed forensic cases in the Western Cape Province of South Africa are lacking.

The effect of NucleoSpin® Forensic Filters on DNA recovery from trace DNA swabs.

Forensic DNA profiling is a globally accepted method for human identification, however, obtaining full DNA profiles from trace DNA can be challenging. The optimal recovery of DNA from trace DNA swabs is therefore crucial. Methods for extracting DNA from swabs often make use of a spin basket combined with a centrifugation step, to enhance the release of cells from the swab prior to DNA extraction.

Towards molecular autopsies: Development of a FFPE tissue DNA extraction workflow.

Sudden unexpected death (SUD) is a devastating event and forms a substantial proportion of the cases investigated at forensic mortuaries each year. Despite post-mortem investigations, the cause of death may remain undetermined. There is potential for these unresolved cases to benefit from retrospective molecular autopsies for investigation into genetic mutations which may have contributed towards death.

A review of the causes and risk factors for sudden unexpected death in the young.

Sudden unexpected death in the young (SUDY) is a tragic event resulting in the fatality of seemingly healthy individuals between the ages of one and 40 years. Whilst studies have been performed on sudden unexpected death in infants, children, and adults respectively, little is known about trends in risk factors and causes of death of SUDY cases. Understanding the factors surrounding these deaths could lead to targeted interventions for at-risk individuals.

Forensic human identification: Investigation into tooth morphotype and DNA extraction methods from teeth.

When a body is decomposed, hard tissues such as teeth may provide the only DNA source for human identification. There is currently no consensus as to the best DNA extraction method, and there is a lack of empirical data regarding tooth morphotype and condition that may impact DNA recovery. Therefore, this study sought to investigate which variables significantly improved DNA concentration, integrity and profiling success.

Massively Parallel Sequencing of 43 Arrhythmia Genes in a Selected SUDI Cohort from Cape Town.

Sudden unexpected death in infants (SUDI) is a devastating event, and unfortunately occurs frequently in developing countries. The emerging molecular autopsy has added value to post-mortem investigations, where genetic variants were able to explain the unexpected demise. Many of these variants have been found in genes involved in arrythmia pathways.

Evaluation of direct PCR for routine DNA profiling of non-decomposed deceased individuals.

Forensic DNA profiling is a standard method used in the attempt to identify deceased individuals. In routine investigations, and if available, the preferred sample type is usually blood. However, this requires the invasive re-opening of the body, days or weeks after the autopsy, which is undesirable in resource-constrained mortuary settings.

A first for forensic genetics in Africa: successful identification of skeletal remains from the marine environment using massively parallel sequencing.

In unrelated circumstances, two young adult males allegedly went missing off the coast of Cape Town, South Africa, within two months of each other. Weeks after the second disappearance, a decomposed human lower limb was recovered from a beach in Cape Town, followed by a washed-up decomposed hand three days later. An item of female clothing was found with the remains, and preliminary analysis of the skeleton indicated a female, leading to confusion regarding the possible identity of the decedent.

Assessment of candidate variants causative of inborn metabolic diseases in SUDI cases in South Africa, and a case report.

Sudden unexpected death in infants (SUDI) is a devastating event, and unfortunately is still a burden in many parts of the world, including in South Africa. Due to the absence of routine testing for inborn metabolic diseases in newborns and in a post-mortem context, little is known about the presence of metabolic diseases in local SUDI cases. The aim of this study was to genotype five candidate variants previously associated with metabolic disorders in a cohort of SUDI cases (n = 169) from Salt River Mortuary, Cape Town.

A retrospective study of death scene investigation practices for sudden unexpected death of infants (SUDI) in Cape Town, South Africa.

A death scene investigation (DSI) forms an integral part of the inquiry into death, particularly for sudden unexpected death of infants (SUDI). Global guidelines exist for DSI, however, previous studies have shown that South Africa does not routinely perform DSI for SUDI cases, nor is there a standard protocol in this regard. This was largely attributed to the large burden of SUDI cases as well as the lack of resources, due to South Africa being a developing country.

Global trends in the extent of death scene investigation performed for sudden and unexpected death of infant (SUDI) cases: A systematic review.

Sudden unexpected death of infants (SUDI) is the rapid and unexpected death of an infant aged less than 1year old. These deaths are referred for a medico-legal investigation to establish cause of death. National and regional protocols for the investigation of SUDI cases have been established in some countries and these typically include a death scene investigation component.

A Systematic Review of Molecular Autopsy Studies in Sudden Infant Death Cases.

Sudden unexpected death is an upsetting event, which can remain unexplained even after post-mortem investigation. Internationally, molecular autopsies have shown to resolve up to 44% of unexplained cases; however, it is currently unclear how many of these were infants. This systematic literature review showed that significantly fewer infant cases were resolved (median: 4%) compared with cohorts of 1 to 45 years old (median: 32%).

Investigation into abandoned neonates admitted to Salt River Forensic Pathology Mortuary, Cape Town.

The high rates of abandoned neonatal remains admitted to Salt River Mortuary (SRM), Cape Town, and the dearth of literature published in South Africa prompted a retrospective assessment of these cases to describe the profile and assess the scope of the post-mortem investigation of abandoned neonates, as well as determine the prosecution rate of abandoned live births. Demographic details, scene information and post-mortem findings were obtained from medico-legal case files concerning non-viable fetuses, stillbirths and concealment of birth cases admitted to SRM between 1 January 2012 and 31 December 2016 (n=249). Despite the majority of the cases being natural deaths, the cause of death frequently remained 'undetermined' in these cases, often due to the presence of decomposition.

DNA barcoding of forensically important flies in the Western Cape, South Africa.

Forensic entomology aids the determination of post mortem interval based on arthropods associated with a deceased body. This relies on the accurate identification of insects that visit the body, particularly first colonisers such as Calliphoridae (Diptera). Traditional species identification though morphological keys can be challenging as immature or closely related specimens can look similar.

A systematic review exploring the relationship between infection and sudden unexpected death between 2000 and 2016: A forensic perspective.

Death due to infectious diseases is a major health concern worldwide. This is of particular concern in developing countries where poor-socio economic status and a lack of healthcare resources contribute to the high burden of disease. In some cases death due to infection can be acute and aggressive, and death may occur without a diagnosis whilst the person is still alive.