DK200301392A - General method for propagating and detecting pathogenic bacteria - Google Patents

Description

PATENT REQUIREMENTS 1. Horizontal method for propagating and detecting pathogenic bacteria in a sample including Salmonella, Listeria, E. coli, Campylobacter, Yersinia, etc. from faeces, food, feed and other products humans or pets can ingest or can enter contact with where other products include but are not limited to feathers, dust, down, wastewater and HACCP samples. which method comprises at least: - a propagation of pathogenic target bacteria in a pre-propagation buffer; and - a transfer of a transfer volume (a sub-volume of the pre-propagation buffer) to a target bacterial selective propagation buffer, which at least contains a propagation buffer. 'agent' that is not tolerated or tolerated worse by the 'non-target' bacteria (following the flora) or other selective pressure on the 'non-target' bacteria such as pH and osmosis,

FEATURED BY - that the pre-propagation medium is heated to above room temperature, preferably above 28 ° C, - that the transfer volume from the pre-amplification to the selective propagation is made after less than 14 hours, and - that the transfer volume is preferably at least 1 ml and at least 1/200 of, respectively. the selective growth medium. 2. Procedure according to Claim 1, characterized in that the concentration of the growth medium in the received selective amplification buffer has a growth media concentration equal to or substantially higher than recommended by the growth media manufacturer or approved methods for the pathogenic target bacterium after the transfer volume has been applied to the selective growth medium. 3. Procedure according to any of the preceding claims characterized by the pre-propagation medium being preheated to a temperature preferably above 30 ° C, more preferably above 32 ° C, most preferably above 34 ° C. 4. Procedure according to Claim 1, characterized in that the pre-propagation medium is preheated to 37 ° C +/- 3 ° C, preferably to 37 ° C. 5. Procedure according to any of the preceding claims characterized in that the transfer of a transfer volume from the pre-amplification to the selective amplification is preferably done after less than 12 hours, more preferably after 6 +/- 4, more preferably after 6 +/- 2 hours, most preferably after 6 hours. . 6. Procedure According to any of the foregoing claims characterized by the transfer of a transfer volume from the pre-amplification to the selective amplification preferably after less than 2 hours. 7. Procedure according to any of the preceding claims characterized in that the transfer volume from the pre-amplification to the selective amplification is preferably at least 2 ml, preferably at least 5 ml, preferably at least 10 ml, preferably at least 20 ml, preferably at least 50 ml, and preferably at least 1/100, preferably at least 1/40, preferably at least 1/20, preferably at least 1/10, preferably at least 1/4 of the selective growth medium. 8. Procedure According to any of the foregoing claims characterized by the method being used to clarify the presence of a single pathogenic bacterium in a tested product. 9. Procedure according to any of the foregoing claims characterized by the pathogenic bacterium being salmonella. 10. Procedure According to CHARACTERISTICS CHARACTERIZED BY SELECTING BIOLINE SUBSTRATE IN SELECTIVE AMPLIFICATION. 11. Method for propagating and detecting pathogenic bacteria according to a sample. any of the foregoing claims FEATURED using a pre-propagation buffer and at least 2 selective growth media. 12. Procedure According to ANY OF THE PRIOR REQUIREMENTS CHARACTERED BY the initial propagation medium being added to growth promoters, resuscitation promoters or selective or partially selective substances such as selenite, tetrathionate, novobiocin, antibiotics, brilliant green or malachite green. 13. Procedure according to any of the foregoing claims characterized by the selective amplification being shortened to 1.2, 4, 6, 8 or 17 hours. 14. Procedure According to any of the foregoing claims FEATURED that the second buffer used in the propagation of target organisms contains tetrathionate, brilliant green, or malachite green. Method of propagating and detecting (if any) pathogenic bacteria in a sample according to. ANY OF THE PRIOR REQUIREMENTS CHARACTERIZED IN THE DETECTION STEP USING AN ANALYTICAL METHOD INCLUDED BUT NOT LIMITED TO: affinity binding techniques such as enzyme immunoassays (ELISA) based on antigen antibody reactions, antigen-antibody reactions involving fluorescence, luminescence, evanescent waves, plasmon resonance, latex agglutination, electrochemical immune detection, immunomagnetic capture techniques, lateral flow techniques, DNA hybridization based on specific sequences of the salmonella DNA molecule, RNA DNA, RNA RNA, Polymerase Chain Reaction (PCR) based on multiplication using specific DNA primers, conductivity measurement method based on change in electrical resistance in particular growth media, microscopy, techniques with micro arrays, CCD camera technique, enzyme immuno-technique based on chromogen, fluorescence, luminescence, radioactive signal generating response, semi-liquid agar and solid agar or combination with outer edges tighter propagation procedures. 16. Method of propagating and detecting (any) pathogenic bacteria in a sample according to. ANY OF THE PRIOR REQUIREMENTS CHARACTERIZED BY using the ELISA test kit from Bioline ApS at the detection stage. 17. A method according to any of the preceding claims, characterized in that the transfer of the transfer volume from the pre-amplification to the selective amplification is carried out automatically or semi-automatically.