Forensic utility of the feline mitochondrial control region – A Dutch perspective

Wesselink, Monique; Bergwerff, Leonie; Hoogmoed, Daniëlle; Kloosterman, A.; Kuiper, Irene

doi:10.1016/j.fsigen.2015.03.004

Cited by 8 publications

(3 citation statements)

References 22 publications

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“…For cat hairs to be used more widely in forensic investigations a more discriminating test is clearly needed. Data from other hypervariable regions [12] and cytochrome b [18] show that discrimination power can be substantially enhanced by sequencing additional parts of the mtDNA molecule.…”

Section: Resultsmentioning

confidence: 99%

Application of a mitochondrial DNA control region frequency database for UK domestic cats

Ottolini

Lall

Sacchini³

et al. 2017

Forensic Science International: Genetics

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DNA variation in 402 bp of the mitochondrial control region flanked by repeat sequences RS2 and RS3 was evaluated by Sanger sequencing in 152 English domestic cats, in order to determine the significance of matching DNA sequences between hairs found with a victim's body and the suspect's pet cat. Whilst 95% of English cats possessed one of the twelve globally widespread mitotypes, four new variants were observed, the most common of which (2% frequency) was shared with the evidential samples. No significant difference in mitotype frequency was seen between 32 individuals from the locality of the crime and 120 additional cats from the rest of England, suggesting a lack of local population structure. However, significant differences were observed in comparison with frequencies in other countries, including the closely neighbouring Netherlands, highlighting the importance of appropriate genetic databases when determining the evidential significance of mitochondrial DNA evidence.

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Section: Resultsmentioning

confidence: 99%

Application of a mitochondrial DNA control region frequency database for UK domestic cats

Ottolini

Lall

Sacchini³

et al. 2017

Forensic Science International: Genetics

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“…Traditionally, the most studied mtDNA region for individualisation purposes is a 402-bp (coordinates: 16,814-206; Fig. 1a) segment within the CR [3,5,[13][14][15][16][17]. This segment is flanked by two distinct repetitive sequence sites, RS2 and RS3 [5], which are reported to display high levels of heteroplasmy, and are generally avoided in analysis [18].…”

Section: Introductionmentioning

confidence: 99%

“…In order to determine the evidential weight of a haplotype, its frequency should be assessed within a reference database large enough to reflect the genetic diversity present in the population [4]. Reference databases of cats sampled in different geographic locations have shown that haplotype frequencies can vary markedly [3,5,13,14,16,17]. Based on analysis of the 402-bp CR segment, twelve 'universal' haplotypes designated A-L have been identified, with the four haplotypes A-D representing 60-70% of cat Fig.…”

Section: Introductionmentioning

confidence: 99%

Defining cat mitogenome variation and accounting for numts via multiplex amplification and Nanopore sequencing

Patterson

Lall

Neumann

et al. 2023

Preprint

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Hair shed by domestic cats is a potentially useful source of forensic evidence. Analysable hair DNA is predominantly mitochondrial, but the recent domestication history of cats means that mtDNA diversity is low. A 402-bp control region segment is usually sequenced, defining only a small number of distinct mitotypes in populations. Previously, we used a long-amplicon approach to sequence whole mitogenomes in a sample of blood DNAs from 119 UK cats, greatly increasing observed diversity and reducing random match probabilities. To exploit this variation for forensic analysis, we here describe a multiplex system that amplifies the cat mitogenome in 60 overlapping amplicons of mean length 360 bp, followed by Nanopore sequencing. Variants detected in multiplex sequence data from hair completely mirror those from long-amplicon data from blood from the same individuals. However, applying the multiplex to matched blood DNA reveals additional sequence variants which derive from the major feline nuclear mitochondrial insertion sequence (numt), which covers 7.9 kb of the 17-kb mitogenome and exists in multiple tandem copies. We use long-amplicon Nanopore sequencing to investigate numt variation in a set of cats, together with an analysis of published genome sequences, and show that numt arrays are variable in both structure and sequence, thus providing a potential source of uncertainty when nuclear DNA predominates in a sample. Forensic application of the test was demonstrated by matching hairs from a cat with skeletal remains from its putative mother, both of which shared a globally common mitotype at the control region. The random match probability (RMP) in this case with the CR 402-bp segment was 0.21 and this decreased to 0.03 when considering the whole mitogenome. The developed multiplex and sequencing approach, when applied to cat hair where nuclear DNA is scarce, can provide a reliable and highly discriminating source of forensic genetic evidence. The confounding effect of numt co-amplification in degraded samples where mixed sequences are observed can be mitigated by variant phasing, and by comparison with numt sequence diversity data, such as those presented here.

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Improvement and automation of a real-time PCR assay for vaginal fluids

Vittori

Giampaoli

Barni³

et al. 2016

Forensic Science International

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Forensic utility of the feline mitochondrial control region – A Dutch perspective

Cited by 8 publications

References 22 publications

Application of a mitochondrial DNA control region frequency database for UK domestic cats

Application of a mitochondrial DNA control region frequency database for UK domestic cats

Defining cat mitogenome variation and accounting for numts via multiplex amplification and Nanopore sequencing

Improvement and automation of a real-time PCR assay for vaginal fluids

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