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Identification and quantification of two species of oyster larvae using real-time PCR⋆

Published online by Cambridge University Press:  15 January 2015

Ana Sánchez ,
Javier Quinteiro ,
Manuel Rey-Méndez ,
Ricardo Isaac Perez-Martín  and
Carmen González Sotelo
Ana Sánchez*
Affiliation:
Instituto de Investigaciones Marinas, Consejo Superior de Investigaciones Científicas (CSIC), Eduardo Cabello 6, Vigo 36208 Pontevedra, Spain
Javier Quinteiro
Affiliation:
Laboratorio de Sistemática Molecular (Unidad Asociada al CSIC), Departamento de Bioquímica y Biología Molecular, CIBUS, Campus Vida, Universidad de Santiago de Compostela, Santiago de Compostela 15782 A Coruña, Spain
Manuel Rey-Méndez
Affiliation:
Laboratorio de Sistemática Molecular (Unidad Asociada al CSIC), Departamento de Bioquímica y Biología Molecular, CIBUS, Campus Vida, Universidad de Santiago de Compostela, Santiago de Compostela 15782 A Coruña, Spain
Ricardo Isaac Perez-Martín
Affiliation:
Instituto de Investigaciones Marinas, Consejo Superior de Investigaciones Científicas (CSIC), Eduardo Cabello 6, Vigo 36208 Pontevedra, Spain
Carmen González Sotelo
Affiliation:
Instituto de Investigaciones Marinas, Consejo Superior de Investigaciones Científicas (CSIC), Eduardo Cabello 6, Vigo 36208 Pontevedra, Spain
*
a Corresponding author: [email protected]
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Abstract

A real-time polymerase chain reaction (PCR) assay was developed for the identification and quantification of two oyster species: Ostrea edulis and Crassostrea gigas. Two sets of primers and TaqMan-MGB probes were designed, based on partial sequences of the 16S rRNA gene. An amplification positive control system was also located in the 18S rRNA gene sequences. Closely related species of oysters and other bivalves, known to co-occur with the target species in European waters, were used to test the assay for cross-reactivity. The assay designed was specific for the target species and no signal or no significant signal was detected for all non-target species tested. The high sensitivity of this method was demonstrated since it is possible to detect just one larva (150–200 μm size) of each species even when it is present with others. Furthermore, this assay provided an acceptable quantification of the number of spiked larvae (1, 10 and 100 larvae) in plankton samples employing a standard curve for larvae.

Keywords

Real-time PCROyster larvae identificationSpecies identification16S rRNA
Type
Research Article
Information
Aquatic Living Resources , Volume 27 , Issue 3-4 , July 2014 , pp. 135 - 145
Copyright
© EDP Sciences, IFREMER, IRD 2015

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Footnotes

⋆

Supporting information is only available in electronic form at www.alr-journal.org.

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