Development of a microplate-based fluorescence immunoassay using quantum dot streptavidin conjugates for enumeration of putative marine bacteria, Alteromonas sp., associated with a benthic harpacticoid copepod

Journal of Microbiological Methods
Erin M BeckmanAlan W Decho

Abstract

Attached bacteria inhabit the surfaces of many marine animals--a process that may play important roles in the survival and transport through aquatic systems. However, efficient detection of these bacteria has been problematic, especially small aquatic animals such as benthic harpacticoid copepod. Quantum dots (QD) have recently emerged as a significant tool in immunofluorescence detection because of their unique properties compared to other fluorescent probes. In the present study, a polyclonal antibody was raised against the Gram-negative marine bacterium, Alteromonas sp. A microplate-based immunofluorescence bioassay using QD strepavidin conjugates was developed for quantifying putative Alteromonas sp. cells located on the surfaces of a marine harpacticoid copepod, Microarthridion littorale. The number of attached Alteromonas sp. was estimated to be 10(2)+/-8 CFU using this method. The QD approach, coupled to a microplate assay can potentially provide an efficient and accurate method for rapidly detecting multiple bacteria species attached to small invertebrate animals because of their unique excitation and emission characteristics.

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Citations

Nov 19, 2010·Bioanalysis·Sonia CentiMarco Mascini
May 5, 2012·Analytical and Bioanalytical Chemistry·N V BeloglazovaI Yu Goryacheva
Mar 15, 2013·Journal of Colloid and Interface Science·Brandon Huey-Ping CheongTuck Wah Ng
Dec 12, 2012·Langmuir : the ACS Journal of Surfaces and Colloids·Xin Ye LiTuck Wah Ng
Dec 5, 2013·Biomacromolecules·Elizabeth M Wurtzler, David Wendell
Sep 7, 2017·Journal of Materials Chemistry. B, Materials for Biology and Medicine·I V MartynenkoY K Gun'ko

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