FN Archimer Export Format
PT J
TI Phytochip: Development of a DNA-microarray for rapid and accurate identification of Pseudo-nitzschia spp and other harmful algal species
BT 
AF NOYER, Charlotte
   ABOT, Anne
   TROUILH, Lidwine
   LEBERRE, Veronique Anton
   DREANNO, Catherine
AS 1:1,2,3,4;2:2,3,4;3:2,3,4;4:2,3,4;5:1;
FF 1:PDG-REM-RDT-LDCM;2:;3:;4:;5:PDG-REM-RDT-LDCM;
C1 IFREMER, Ctr Bretagne, Lab Detect Capteurs & Mesures, F-29280 Plouzane, France.
   Univ Toulouse, INSA, UPS, INP,LISBP, F-31077 Toulouse, France.
   INRA, UMR792, Ingn Syst Biol & Proc, F-31400 Toulouse, France.
   CNRS, UMR5504, F-31400 Toulouse, France.
C2 IFREMER, FRANCE
   UNIV TOULOUSE, FRANCE
   INRA, FRANCE
   CNRS, FRANCE
SI BREST
SE PDG-REM-RDT-LDCM
IN WOS Ifremer jusqu'en 2018
   copubli-france
   copubli-p187
   copubli-univ-france
IF 1.857
TC 19
UR https://archimer.ifremer.fr/doc/00254/36554/42346.pdf
LA English
DT Article
DE ;HAB;Pseudo-nitzschia;Microarray;Ribosomal oligonucleotide probes;Environmental monitoring
AB Detection of harmful algal blooms has become a challenging concern because of to the direct impacts on public health and economy. The identification of toxic dinoflagellates and diatoms in monitoring programs requires an extensive taxonomic expertise and is time consuming. Advances in molecular biology have allowed the development of new approaches, more rapid, accurate and cost-effective for detecting these microorganisms. In this context, we developed a new DNA microarray (called, Phytochip) for the simultaneous detection of multiple HAB species with a particular emphasis on Pseudo-nitzschia species. Oligonucleotide probes were designed along the rRNA operon. After DNA extraction, the target rDNA genes were amplified and labeled using an asymmetric PCR; then, the amplicons were hybridized to the oligonucleotide probes present on the chips. The total assay from sea water sampling to data acquisition can be performed within a working day. Specificity and sensitivity were assessed by using monoclonal cultures, mixtures of species and field samples spiked with a known amount of cultured cells. The Phytochip with its 81 validated oligonucleotide probes was able to detect 12 species of Pseudo-nitzschia and 11 species of dinoflagellates among which were 3 species of Karenia and 3 species of Alexandrium. The Phytochip was applied to environmental samples already characterized by light microscopy and cloned into DNA libraries. The hybridizations on the Phytochip were in good agreement with the sequences retrieved from the clone libraries and the microscopic observations. The Phytochip enables a reliable multiplex detection of phytoplankton and can assist a water quality monitoring program as well as more general ecological research.
PY 2015
PD MAY
SO Journal Of Microbiological Methods
SN 0167-7012
PU Elsevier Science Bv
VL 112
UT 000353426900011
BP 55
EP 66
DI 10.1016/j.mimet.2015.03.002
ID 36554
ER

EF