DIAGNOSTIC KIT AND METHOD FOR THE SIMULTANEOUS DETECTION OF A COMBINATION OF MULTIPLE GRAM-POSITIVE BACTERIA AND/OR GRAM-NEGATIVE BACTERIA

摘要

A method for the simultaneous detection of a combination of multiple Gram-positive bacteria and/or Gram-negative bacteria in a sample, the method comprising: providing DNA extracted from the sample; preparing a reaction mixture containing the DNA, one or more pairs of oligonucleotide primers having a sequence selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2, SEQ ID NO: 4 and SEQ ID NO: 5, SEQ ID NO: 7 and SEQ ID NO: 8, SEQ ID NO: 10 and SEQ ID NO: 11, SEQ ID NO: 13 and SEQ ID NO: 14, SEQ ID NO: 16 and SEQ ID NO: 17 and combinations of same, and one or more oligonucleotide probes having a sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 6, SEQ ID NO: 9, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 18 and combinations of same; amplifying the DNA included in the reaction mixture, by means real-time polymerase chain reaction; and simultaneously determining the presence or absence of Gram-positive bacteria and/or Gram-negative bacteria in the sample. The method can be used to identify any combination of three species of Gram-positive bacteria (Listeria ssp., Listeria monocytogenes and Staphylococcus aureus) and three species of Gram-negative bacteria (Escherichia coli spp.,Escherichia coli O157:H7 and Salmonella spp.).