A method for extracting DNA from hard tissues for use in forensic identification

Samsuwan, Jarunya; Somboonchokepisal, Thanutham; Akaraputtiporn, Thunyathorn; Srimuang, Tunwarut; Phuengsukdaeng, Phuris; Suwannarat, Aunchulee; Mutirangura, Apiwat; Kitkumthorn, Nakarin

doi:10.3892/br.2018.1148

Cited by 11 publications

(11 citation statements)

References 23 publications

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“…Six of them were proven weakly ASFV‐positive by PCR, they were all detected in the infected area between October 2019 and March 2020. For these six weakly ASFV‐positive (Ct > 34) bones found in 2020, based on the date of discovery, climatic conditions and macroscopic appearance, the post‐mortem interval was estimated at > 6 months, according to Samsuwan and colleagues (2018). Further, the ASFV EURL (INIA, Madrid, Spain) was unable to isolate infectious virus from these samples.…”

Section: Assessmentmentioning

confidence: 99%

Epidemiological analysis of African swine fever in the European Union (September 2019 to August 2020)

Daniel¹,

Gerbier²,

Schmidt³

et al. 2021

EFS2

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An update on the African swine fever ( ASF ) situation in the 10 affected Member States ( MS ) in the EU and in two neighbouring countries from the 1 September 2019 until the 31 August 2020 is provided. The dynamics of the proportions of PCR ‐ and ELISA ‐positive samples since the first ASF detection in the country were provided and seasonal patterns were investigated. The impact of the ASF epidemic on the annual numbers of hunted wild boar in each affected MS was investigated. To evaluate differences in the extent of spread of ASF in the wild boar populations, the number of notifications that could be classified as secondary cases to a single source was calculated for each affected MS and compared for the earliest and latest year of the epidemic in the country. To evaluate possible risk factors for the occurrence of ASFV in wild boar or domestic pigs, a literature review was performed. Risk factors for the occurrence of ASF in wild boar in Romanian hunting grounds in 2019 were identified with a generalised linear model. The probability to find at least one PCR ‐confirmed ASF case in wild boar in a hunting ground in Romania was driven by environmental factors, wild boar abundance and the density of backyard pigs in the hunting ground area, while hunting‐related variables were not retained in the final model. Finally, measures implemented in white zones ( ASF ‐free zones that are geographically adjacent to an area where ASF is present in wild boar) to prevent further spread of ASF were analysed with a spatially, explicit stochastic individual‐based model. To be effective, the wild boar population in the white zone would need to be drastically reduced before ASF arrives at the zone and it must be wide enough. To achieve the necessary pre‐emptive culling targets of wild boar in the white zone, at the start of the establishment, the white zone should be placed sufficiently far from the affected area, considering the speed of the natural spread of the disease. This spread is faster in denser wild boar populations. After a focal ASF introduction, the white zone is always close to the infection hence pre‐emptive culling measures in the white zone must be completed in short term, i.e. in a few months.

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Section: Assessmentmentioning

confidence: 99%

Epidemiological analysis of African swine fever in the European Union (September 2019 to August 2020)

Daniel¹,

Gerbier²,

Schmidt³

et al. 2021

EFS2

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“…Molecular biology approaches have been used to evaluate the relationships between environmental insults and DNA degradation [ 10 , 11 , 12 , 13 ]. Because DNA is often highly fragmented [ 14 , 15 ], mitochondrial DNA could be more useful than nuclear DNA [ 15 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Parameters for Estimating the Postmortem Interval of Skeletal Remains Using Bovine Femurs: A Pilot Study

et al. 2020

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The postmortem interval (PMI) of victims is a key parameter in criminal investigations. However, effective methods for estimating the PMI of skeletal remains have not been established because it is determined by various factors, including environmental conditions. To identify effective parameters for estimating the PMI of skeletal remains, we investigated the change in bone focusing on the amount of DNA, element concentrations, and bone density that occurred in the bone samples of bovine femurs, each maintained under one of five simulated environmental conditions (seawater, freshwater, underground, outdoors, and indoors) for 1 year. The amount of extracted mitochondrial DNA (mtDNA; 404 bp fragment) decreased over time, and significant DNA degradation (p < 0.01), as estimated by a comparison with amplification results for a shorter fragment (128 bp), was detected between 1 month and 3 months. Eleven of 30 elements were detected in samples by inductively coupled plasma optical emission spectrometry, and Na and Ba showed significant quantitative differences in terms of environmental conditions and time (p < 0.01). This preliminary study suggests that the level of DNA degradation determined by real-time polymerase chain reaction and element concentrations determined by inductively coupled plasma optical emission may be useful indices for estimating the PMI of victims under a wide range of environmental conditions. However, this study is a limited experimental research and not applicable to forensic cases as it is. Further studies of human bone with longer observation periods are required to verify these findings and to establish effective methods for PMI estimation.

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“…The ability to generate genetic data from highly compromised samples, as often encountered in wildlife forensic science, is essential such that the resulting data are not prone to false assignment [52][53][54][55][56] . Ideal amplified fragment size proposed for SNaPshot assay ranges from 60 to 100 bp 57 .…”

Section: Discussionmentioning

confidence: 99%

Forensic validation of a panel of 12 SNPs for identification of Mongolian wolf and dog

Jiang

et al. 2020

Sci Rep

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Wolf (Canis lupus) is a species included in appendices of citeS and is often encountered in cases of alleged poaching and trafficking of their products. When such crimes are suspected, those involved may attempt to evade legal action by claiming that the animals involved are domestic dogs (C. l. familiaris). To respond effectively to such claims, law enforcement agencies require reliable and robust methods to distinguish wolves from dogs. Reported molecular genetic methods are either unreliable (mitogenome sequence based), or operationally cumbersome and require much DNA (un-multiplexed microsatellites), or financially expensive (genome wide SNP genotyping). We report on the validation of a panel of 12 ancestral informative single nucleotide polymorphism (SNP) markers for discriminating wolves from dogs. A Snapshot multiplex genotyping system was developed for the panel, and 97 Mongolian wolves (C. l. chanco) and 108 domestic dogs were used for validation. Results showed this panel had high genotyping success (0.991), reproducibility (1.00) and origin assignment accuracy (0.97 ± 0.05 for dogs and 1.00 ± 0.03 for wolves). Species-specificity testing suggested strong tolerance to DNA contamination across species, except for Canidae. The minimum DNA required for reliable genotyping was 6.25 pg/μl. The method and established gene frequency database are available to support identification of wolves and dogs by law enforcement agencies. Grey wolf (Canis lupus) was once abundant over much of Eurasia and North America. However, some regional populations have suffered a severe decline in numbers because of habitat destruction and human interference 1-3 , poaching 4,5 , and human-wolf conflicts 6,7. Although the global population is stable 8 , the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) list grey wolf in Appendix I (populations of Bhutan, India, Nepal and Pakistan) and Appendix II (other populations) 9. The objective of these listings is to restrict international trade in wolves and wolf body parts. The species is also protected by national laws of most range countries. Culturally, wolf bodies and body parts are often believed to have magical powers 10 and medicinal functions 11-15. Such unfounded superstitions as well as human-wolf conflicts are factors that drive poaching and trafficking of wolves and body parts in Eurasian countries. For example, media reports from China include at least 35 trafficking cases uncovered by China customs from 2010 to 2018. Seizures included hundreds of skins, carcasses, mounted specimens, and more than a thousand carpal bones and canines being trafficked from Mongolia, Russia, Kazakhstan, and Kyrgyzstan. In attempts to avoid penalties, traders often claimed that the bones and canines were those of large dogs and were sold as counterfeits of wolf body parts. Prosecution of these cases is dependent on definitive subspecies identification. There are many morphological differences between wolves and dogs. The most apparent differentiation is base...

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A method for extracting DNA from hard tissues for use in forensic identification

Cited by 11 publications

References 23 publications

Epidemiological analysis of African swine fever in the European Union (September 2019 to August 2020)

Epidemiological analysis of African swine fever in the European Union (September 2019 to August 2020)

Evaluation of Parameters for Estimating the Postmortem Interval of Skeletal Remains Using Bovine Femurs: A Pilot Study

Forensic validation of a panel of 12 SNPs for identification of Mongolian wolf and dog

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