Species-Specific
Oligonucleotides
(SSOs)
as
a
forensic DNA test [HD
Marshall,
KA Johnstone, & SM Carr (2007) Forensic Sci Int'l,167, 1-7]
The
concept of allele-specific oligonucleotides
(ASOs) can be extended to
species-specific
oligonucleotides (SSOs).
The DNA sequences of the
Cytochrome Oxidase I gene
are known
for many species of scallops, including two found in the Northwest
Atlantic, Sea Scallops (Placopecten
magellanicus) and Icelandic Scallops (Chlamys icelandica). The
multiplex
test shown here combines an anchor
primer for a DNA sequence
identical
in
both species (ScallopR2)
at
position 932 with primers specific for either species (PmaCOIF1
and CisCOIF2),
which
are differentiated at positions 313 and 473, respectively. The PCR amplification product
from Placopecten is
thus expected to be
932 - 313 = 619bpin length,
versus a
shorter 932 - 473 = 459bp in Chlamys. That is, the size of the
fragment indicates species identity directly.
In
"The Case of
the Scurrilous Scallops", a fisherman had a load of
scallops that he claimed were from the open fishery for Islandic
scallops (Chlamys).
Enforcement officers suspected they were instead from the closed
fishery for Sea scallops (Placopecten).
Since
a
small
proportion of bycatch from the closed fishery might be
considered acceptable, the legal question was: What fraction of
the
total catch was from the prohibited species? DNA was extracted from
individual
scallops and amplified in the multiplex SSO test with the three primers. Of
the 80 scallops in the test above, all but eight (blue arrows)
show
the larger DNA fragment,
which indicates that 90% are Placopecten.
In
the
complete series of more than 900 scallops from two vessels,
almost two-thirds were Placopecten.
This
resulted
in conviction and fine.
The multiplex SSO
test
provides
a
rapid, direct means of forensic identification of large
population sample series, without the necessity of secondary DNA sequencing, RFLP mapping, or
fingerprinting,
and can be adapted to other loci and species.