US Government as Represented by the Secretary of the Army

Sheila J Wood

Barbara J Mann

Robert J Sydiskis

In one aspect of the invention there is described a method of detecting the presence of toxins, which may traverse bacterial membranes in some manner, in environmental samples using an indicator bacteria such as E. coli K-12 and a fluorescent signal. The method includes:

Method and kit for rapid detection of toxins and bacteria

A method of identifying pathogens, comprising the steps of forming a micrhere having a fluorescent compound attached thereto as a solid matrix for the fluorescent compound. A single strand of DNA is immobilized on the matrix to form a probe, and prior to that a short complementary central piece is formed with a fluorescent compound labelled on one end thereof which joins the central piece to the probe. The two strands bind to form a microsphere/probe complex which is subjected to contact with a complementary strand of an unknown sample to displace the centrally attached probe appendage to provide an increase in fluorescent signal indicative of the emission maxima of the dye in the microsphere to verify the similarity of the unknown sample with known members of the complex. It will be noted that transfer of resonant energy between the complex and the unknown sample identifies the presence of an antigen an adhesin or an antibody, the transfer occurring by use of two fluorescent compounds, one of which absorbs energy at substantially the same wavelength as the other emits energy. Similarly, the presence of a particular sequence of DNA in the unknown sample is determined by fabricating the complementary strand, lysing the organism, splitting the strand of DNA and mixing the resulting probe with the unknown sample, whereby junction of the two strands is affirmed by a chemical signal from the fluorescent dye.

Comprehensive identification scheme for pathogens

A novel laboratory method and a kit for rapid detection and identification of Yersinia bacteria using the polymerase chain reaction with specific primers and specific restriction enzymes to differentiate among Yersinia at the species level and to separate Yersinia from other species of bacteria.

Detection of Yersinia using the polymerase chain reaction

A microorganism detection system provides initial warning, confirmation of dentity, and recognition of pathogenic factors in microorganisms from environmental samples. The method and apparatus of the invention uses different sized antibody coated microspheres which react with unknown antigens, are sized by electronic volume sizing, and are sorted by size. The sized particles are quantitated in addition to being sized. The microsphere sizes indicate the presence of specific microorganism groups. The samples can be further analyzed using fluorescent microspheres which agglutinate with the sized microspheres. The presence of specific microorganisms is indicated by a change in the fluorescence of the sample.

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Edgewood, MD, United States of America

Sheila J Wood