title: Molecular detection of a potentially toxic diatom species.
abstract: 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.
authors.affiliation: Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, Camerino 62032, Italy. bidhan.dhar@gmail.com.
authors.forename: Bidhan Chandra
authors.initials: BC
authors.lastname: Dhar
authors.affiliation: Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, Camerino 62032, Italy. lucia.cimarelli@studenti.unicam.it.
authors.forename: Lucia
authors.initials: L
authors.lastname: Cimarelli
authors.affiliation: Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, Camerino 62032, Italy. kumar.saurabhsingh@unicam.it.
authors.forename: Kumar Saurabh
authors.initials: KS
authors.lastname: Singh
authors.affiliation: Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, Camerino 62032, Italy. letizia.brandi@unicam.it.
authors.forename: Letizia
authors.initials: L
authors.lastname: Brandi
authors.affiliation: Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, Camerino 62032, Italy. anna.brandi@unicam.it.
authors.forename: Anna
authors.initials: A
authors.lastname: Brandi
authors.affiliation: Environmental, Quality and Fishfarm Unit, Environment & Primary Prevention Department, Istituto Superiore di Sanità, Viale Regina Elena, Rome 299, Italy. camilla.puccinelli@iss.it.
authors.forename: Camilla
authors.initials: C
authors.lastname: Puccinelli
authors.affiliation: Environmental, Quality and Fishfarm Unit, Environment & Primary Prevention Department, Istituto Superiore di Sanità, Viale Regina Elena, Rome 299, Italy. stefania.marcheggiani@iss.it.
authors.forename: Stefania
authors.initials: S
authors.lastname: Marcheggiani
authors.affiliation: Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, Camerino 62032, Italy. roberto.spurio@unicam.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Spurio
mesh_terms: D017377:Diatoms / Q000302:isolation & purification*; D007558:Italy; D008522:Mediterranean Sea; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes
chemical_list: D015345:Oligonucleotide Probes