The selling and importing of puffer fish species and their products was banned in Thailand in 2002, because of possible neurotoxic effects. However, the sale of their flesh is still happening in Thai markets. Standard methods for toxin quantification (HPLC and LC-MS) have significant limitations, therefore a lateral flow, immuno-chromatographic test (TTX-IC) was developed as a tool for rapid detection of toxin. A total of 750 puffer fishes (387 Lagocephalus lunaris(LL), and 363 Lagocephalus spadiceus (LS)) and 100 edible fishes were caught in Thailand from June 2011-February 2012. Screening of TTX from their flesh by TTX-IC revealed that 69 samples (17.8%) of LL possessed TTX at dangerous levels but LS and edible fishes did not. A selected 339 samples were quantified by LC-MS/MS, showing 50 LL possessed TTX at dangerous levels. Comparison of results with LC-MS/MS showed the TTX-IC to have 94.0% sensitivity and 92.4% specificity. The TTX-IC will be a useful tool for TTX screening of a large number of samples, reducing the testing required by LC-MS/MS, thus reducing costs. All positive cases found should be confirmed by standard methods.
This study demonstrated the potential application of antibody-conjugated Rubpy dye-doped silica nanoparticles for immunofluorescence microscopic detection ofVibrio choleraeO1. The particle synthesis of 20X of the original ratio was accomplished yielding spherical nanoparticles with an average size of45±3 nm. The nanoparticles were carboxyl functionalized and then conjugated with either monoclonal antibody or polyclonal antibody againstV. choleraeO1. The antibody-conjugated nanoparticles were tested with two target bacteria and three challenge strains. The result showed that monoclonal antibody-conjugated Rubpy dye-doped silica nanoparticles could be effectively used as signal amplification to detectV. choleraeO1 under a fluorescence microscope. Their extremely strong fluorescence signal also enables the detection of a single cell bacterium.
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