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2015 ; 12
(5
): 4921-41
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Molecular detection of a potentially toxic diatom species
#MMPMID25955528
Dhar BC
; Cimarelli L
; Singh KS
; Brandi L
; Brandi A
; Puccinelli C
; Marcheggiani S
; Spurio R
Int J Environ Res Public Health
2015[May]; 12
(5
): 4921-41
PMID25955528
show ga
A few diatom species produce toxins that affect human and animal health. Among
these, members of the Pseudo-nitzschia genus were the first diatoms unambiguously
identified as producer of domoic acid, a neurotoxin affecting molluscan
shell-fish, birds, marine mammals, and humans. Evidence exists indicating the
involvement of another diatom genus, Amphora, as a potential producer of domoic
acid. We present a strategy for the detection of the diatom species Amphora
coffeaeformis based on the development of species-specific oligonucleotide probes
and their application in microarray hybridization experiments. This approach is
based on the use of two marker genes highly conserved in all diatoms, but endowed
with sufficient genetic divergence to discriminate diatoms at the species level.
A region of approximately 450 bp of these previously unexplored marker genes,
coding for elongation factor 1-a (eEF1-a) and silicic acid transporter (SIT), was
used to design oligonucleotide probes that were tested for specificity in
combination with the corresponding fluorescently labeled DNA targets. The results
presented in this work suggest a possible use of this DNA chip technology for the
selective detection of A. coffeaeformis in environmental settings where the
presence of this potential toxin producer may represent a threat to human and
animal health. In addition, the same basic approach can be adapted to a wider
range of diatoms for the simultaneous detection of microorganisms used as
biomarkers of different water quality levels.