Sequence polymorphism-based identification and quantification of Vibrio nigripulchritudo at the species and subspecies level targeting an emerging pathogen for cultured shrimp in New Caledonia.
In a previous study, we demonstrated the existence of an emerging cluster of Vibrio nigripulchritudo that proved to be associated with shrimp mortality events in New Caledonia. Using sequence polymorphisms evidenced in this previous MultiLocus Sequence Typing study, we developed two new quantitative PCR assays permitting the detection and quantification of V. nigripulchritudo at the genospecies level using SYBR Green I chemistry and at the emerging cluster level using Fluorescence Resonance Energy Transfer technology with hybridization probes. The use of this molecular diagnostic tool evidenced the colonization of the shrimp pond ecosystem by the pathogenic cluster at least at the onset of the disease. This new tool will allow better investigation of the dynamics of this bacterial pathogen in the shrimp farm ecosystem.
Goarant
Cyrille
C
Laboratoire de Recherche en Bactériologie, Institut Pasteur de Nouvelle Calédonie, Noumea, New Caledonia. cgoarant@ifremer.fr
Reynaud
Yann
Y
Ansquer
Dominique
D
de Decker
Sophie
S
Merien
Fabrice
F
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D052160:Benzothiazoles; D015169:Colony Count, Microbial / Q000379:methods*; D027081:DNA Gyrase / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D003959:Diamines; D031541:Fluorescence Resonance Energy Transfer; D008969:Molecular Sequence Data; D009510:New Caledonia; D009693:Nucleic Acid Hybridization; D009930:Organic Chemicals / Q000378:metabolism; D033561:Penaeidae / Q000382:microbiology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D011804:Quinolines; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D014733:Vibrio / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*
D001426:Bacterial Proteins; D052160:Benzothiazoles; D004269:DNA, Bacterial; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D027081:DNA Gyrase