Pyrosequencing: Current forensic methodology and future applications—a review

The recent development of small, single‐amplicon‐based benchtop systems for pyrosequencing has opened up a host of novel procedures for applications in forensic science. Pyrosequencing is a sequencing by synthesis technique, based on chemiluminescent inorganic pyrophosphate detection. This review ex...

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Bibliographic Details
Published in:Electrophoresis 2023-01, Vol.44 (1-2), p.298-312
Main Authors: Ghemrawi, Mirna, Tejero, Nicole Fernandez, Duncan, George, McCord, Bruce
Format: Article
Language:English
Subjects:
age
body fluid ID
Body fluids
Body size
Chemiluminescence
Design factors
Drug abuse
epigenetics
Forensic Genetics
Forensic Medicine
Forensic science
Forensic Sciences
forensics
Genomics
High-Throughput Nucleotide Sequencing - methods
phenotyping
pyrosequencing
species ID
Workflow
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Summary:The recent development of small, single‐amplicon‐based benchtop systems for pyrosequencing has opened up a host of novel procedures for applications in forensic science. Pyrosequencing is a sequencing by synthesis technique, based on chemiluminescent inorganic pyrophosphate detection. This review explains the pyrosequencing workflow and illustrates the step‐by‐step chemistry, followed by a description of the assay design and factors to keep in mind for an exemplary assay. Existing and potential forensic applications are highlighted using this technology. Current applications include identifying species, identifying bodily fluids, and determining smoking status. We also review progress in potential applications for the future, including research on distinguishing monozygotic twins, detecting alcohol and drug abuse, and other phenotypic characteristics such as diet and body mass index. Overall, the versatility of the pyrosequencing technologies renders it a useful tool in forensic genomics.
ISSN:0173-0835
1522-2683
DOI:10.1002/elps.202200177