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Bogush S. S., Bulychiova Yu. A., Gavrilova Ye. M., Yegorov A. M.

A Model for the Enzyme-Linked Immunosorbent Determination of Cyclosporin A in Whole Blood

Abstract

Bioluminescent assay of total bacterial contamination (total number of mesophilic aerobic and facultative anaerobic microorganisms) of raw processed meat, based on determination of intracellular bacterial ATP was developed. For elimination of non-bacterial ATP the sample was treated with mixture of detergent and protease followed by filtration through bacterial membrane filter. For simplification of the assay the special luminometric microcuvette FiltravetteTM was used. The bottom of FiltravetteTM is made of bacterial membrane filter. This design allows to concentrate bacteria cells, release intracellular ATP and measure bioluminescent signal in the same cuvette. Combined application of the highly sensitive ATP-reagent based on recombinant firefly luciferase which was developed in our laboratory, and FiltravetteTM allows to detect bacterial contamination with detection limit 103 CFU per/g meat. The duration of the assay is about 100 min while the standard Plate Count takes 48 h. A good correlation between bioluminescent assay and the standard Plate Count was observed (R=0,99).
Moscow University Chemistry Bulletin.
2002, Vol. 43, No. 6, P. 392
   

Copyright (C) Chemistry Dept., Moscow State University, 2002
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