Pyrosequencing: Current forensic methodology and future applications—a review

Ghemrawi, Mirna; Tejero, Nicole Fernández; Duncan, George; McCord, Bruce

doi:10.1002/elps.202200177

Cited by 10 publications

(2 citation statements)

References 136 publications

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“…They include pyrosequencing [15,25,[31][32][33], massive parallel sequencing (MPS) [34][35][36][37], SNaPshot assays [38,39], and EpiTYPER [13,28,40,41]. It is important to point out that methylation profiles obtained with these different sequencing/typing methods provide largely comparable results [42].…”

Section: Discussionmentioning

confidence: 99%

An ELOVL2-Based Epigenetic Clock for Forensic Age Prediction: A Systematic Review

Paparazzo

Lagani

Geracitano

et al. 2023

IJMS

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The prediction of chronological age from methylation-based biomarkers represents one of the most promising applications in the field of forensic sciences. Age-prediction models developed so far are not easily applicable for forensic caseworkers. Among the several attempts to pursue this objective, the formulation of single-locus models might represent a good strategy. The present work aimed to develop an accurate single-locus model for age prediction exploiting ELOVL2, a gene for which epigenetic alterations are most highly correlated with age. We carried out a systematic review of different published pyrosequencing datasets in which methylation of the ELOVL2 promoter was analysed to formulate age prediction models. Nine of these, with available datasets involving 2298 participants, were selected. We found that irrespective of which model was adopted, a very strong relationship between ELOVL2 methylation levels and age exists. In particular, the model giving the best age-prediction accuracy was the gradient boosting regressor with a prediction error of about 5.5 years. The findings reported here strongly support the use of ELOVL2 for the formulation of a single-locus epigenetic model, but the inclusion of additional, non-redundant markers is a fundamental requirement to apply a molecular model to forensic applications with more robust results.

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Section: Discussionmentioning

confidence: 99%

An ELOVL2-Based Epigenetic Clock for Forensic Age Prediction: A Systematic Review

Paparazzo

Lagani

Geracitano

et al. 2023

IJMS

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“…Pyrosequencing is a sequencing‐by‐synthesis technique that results in a signal as nucleotides are added to the DNA strand. As each nucleotide is incorporated, a pyrophosphate is released and produces a chemiluminescent signal [12]. Pyrosequencing poses a great advantage in forensics because it is cost‐effective and rapid.…”

Section: Introductionmentioning

confidence: 99%

An examination of differences in epigenetic methylation of saliva type samples based on collection method

Ghemrawi,

Fernandez‐Tejero,

Vaquero

et al. 2024

Electrophoresis

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In the context of forensic casework, it is imperative to both establish a DNA profile from biological specimens and accurately identify the specific bodily fluid source. To achieve this, DNA methylation markers have been developed for the differentiation of blood, semen, vaginal epithelial secretions, and saliva samples. Saliva, alternatively referred to as oral fluid, is recognized for its heterogeneous cellular composition, characterized by a mixture of epithelial, leukocytic, and bacterial cells. Consequently, our research has revealed variations in methylation percentages that correlate with the method employed for collecting saliva samples. To investigate these concepts, we scrutinized four CpG markers situated within or in proximity to the BCAS4, SLC12A8, SOX2OT, and FAM43A genes. Subsequently, we designed primers based on bioinformatically transformed reference sequences for these markers and rigorously assessed their quality by examining dimer and hairpin formation, melting temperature, and specificity. These loci were identified as saliva markers based on either buccal swabs or spit collection. Yet, there has been minimal or no research conducted to explore the variations in methylation between different collection methods. For this study, buccal, lip, tongue, spit, and nasal swabs were collected from 20 individuals (N = 100). Mock forensic samples, which include chewing gum (N = 10) and cigarettes (N = 10), were also tested. DNA was extracted, bisulfite converted, then amplified using in‐house designed assays, and pyrosequenced. The methylation levels were compared to other body fluids (semen, blood, vaginal epithelia, and menstrual blood [N = 32]). A total of 608 pyrosequencing results demonstrated that sampling location and collection method can greatly influence the level of methylation, highlighting the importance of examining multiple collection/deposition methods for body fluids when developing epigenetic markers.

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A novel long noncoding RNA located on the antisense strand of MAL promotes the invasion and metastasis of oral squamous cell carcinoma

Deng

Wang

Zhang

et al. 2023

Archives of Oral Biology

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Pyrosequencing: Current forensic methodology and future applications—a review

Cited by 10 publications

References 136 publications

An ELOVL2-Based Epigenetic Clock for Forensic Age Prediction: A Systematic Review

An ELOVL2-Based Epigenetic Clock for Forensic Age Prediction: A Systematic Review

An examination of differences in epigenetic methylation of saliva type samples based on collection method

A novel long noncoding RNA located on the antisense strand of MAL promotes the invasion and metastasis of oral squamous cell carcinoma

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