Highlights
- •A novel primer set to identify python species in forensic inquiries.
- •Phylogenetic reconstruction accurately identifies species with statistical support.
- •Primer pair shows high sensitivity, amplifying 30 fg or 9 mitochondrial DNA copies.
- •The assay has demonstrated suitability to samples of poor quality.
- •Validation tests demonstrate this assay as suitable for forensic investigations.
Abstract
Python snake species are often encountered in illegal activities and the question
of species identity can be pertinent to such criminal investigations. Morphological
identification of species of pythons can be confounded by many issues and molecular
examination by DNA analysis can provide an alternative and objective means of identification.
Our paper reports on the development and validation of a PCR primer pair that amplifies
a segment of the mitochondrial cytochrome b gene that has been suggested previously as a good candidate locus for differentiating
python species. We used this DNA region to perform species identification of pythons,
even when the template DNA was of poor quality, as might be the case with forensic
evidentiary items. Validation tests are presented to demonstrate the characteristics
of the assay. Tests involved the cross-species amplification of this marker in non-target
species, minimum amount of DNA template required, effects of degradation on product
amplification and a blind trial to simulate a casework scenario that provided 100%
correct identity. Our results demonstrate that this assay performs reliably and robustly
on pythons and can be applied directly to forensic investigations where the presence
of a species of python is in question.
Keywords
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Article info
Publication history
Published online: December 11, 2014
Accepted:
December 9,
2014
Received in revised form:
November 28,
2014
Received:
October 24,
2014
Identification
Copyright
© 2014 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
