Development of a non-enzyme based capillary dipstick assay for the detection and purification of environmental pathogens

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University of Hawaii at Manoa

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Novel mAbs were produced with a wide range of anti-Dickeya binding reactivities. A mAb line produced from the first fusion generated a BOX-PCR type B,C and D binding antibody (MAb Pine-1). A second mAb line had broad spectrum specificity that reacts to BOX-PCR type A through E and most of the Dickeya reference strains (MAb Pine-2). Phylogenetic analysis based on gyr-B gene expression and BOX-PCR types are consistent with the mAb reactivities. Also a field dipstick assay was developed for the detection of live environmental pathogens that is able concentrate the organism for further analysis and was able to detect S. enterica serotype Typhimurium at 105 CFU/ml, which is within the infective dose of most Salmonella species. The dipstick is able to isolate and purify S. enterica serotype Typhimurium from a mixed culture of 100:1 S. marcescens to S. enterica serotype Typhimurium, with a final purity of greater than 80% determined by colony counts, which is culturable to streak for identification.

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Theses for the degree of Master of Science (University of Hawaii at Manoa). Microbiology.

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