Systems and methods for microbe identification (ID) in the context of phenotypic antimicrobial susceptibility testing (AST). Approaches for rapidly identifying polymicrobial samples are introduced that following perform one or more ID methods, such as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF), PCR, or DNA hybridization, following sample incubation in two or more different media, including at least one selective media. Parallel ID and AST workflows are provided that may reduce the time from sample to AST result.

The present invention features general-purpose microbiological growth media capable of supporting growth of microorganisms on membranes. The media contain casein digest, soybean digest, animal tissue digest, yeast extract, dextrose, a phosphate buffer, hemin, and L-cystine. The invention features an all-purpose microbiological growth media that can support the growth of anaerobes, molds, injured spores, and general aerobic bacteria to a greater extent than other media.

Selective enrichment media and methods for selectively growing and detecting Salmonella spp. and/or Shiga toxin-producing E. coli. The media may comprise a carbon and nitrogen source, an inorganic salt, a fermentable sugar, one or more selective agents, and an efflux pump inhibitor. Various selective agents include sulfa drugs, surfactants, aminocoumarins, cycloheximide, supravital stains, ascorbic acid, bromobenzoic acid, myricetin, nitrofurantoin, rifamycins, polyketides, and oxazolidinones. Various efflux pump inhibitors include arylpiperazines, such as 1-(1-naphthylmethyl)piperazine, and quinoline derivatives, such as 4-chloroquinoline. Methods of selectively growing and detecting Salmonella and/or Shiga toxin-producing E. coli are provided.

The present invention relates to a method of directly detecting, using a mass-spectrometry method, whether a microorganism contained in a sample is resistant to antibiotics, and a kit for detection used therewith. More particularly, the present invention relates to a method and kit for directly detecting an antibiotic hydrolase secreted by a microorganism resistant to antibiotics, thereby directly determining whether the microorganism is resistant to antibiotics. According to the present invention, it is possible to very simply and immediately confirm whether a specific strain is resistant to antibiotics in the field. In particular, a complicated pretreatment process such as proteolysis is not performed, and a complicated identification process of calibrating and then combining the obtained results is not performed. Accordingly, it is possible to realize a method of easily confirming whether antibiotic resistance occurs in just a dozen minutes, compared to a conventional technology in which it takes several days to confirm whether antibiotic resistance occurs, and a simple diagnostic kit used therewith.

A bacterium belonging to the genus Pseudomonas such as Pseudomonas aeruginosa is specifically and quantitatively detected using a selection medium for a bacterium belonging to the genus Pseudomonas characterized by containing, in a basal medium, at least one third-generation cephalosporin and at least one antibiotic selected from the group consisting of a first-generation cephalosporin and a glycopeptide-based antibiotic and by a method for detecting a bacterium belonging to the genus Pseudomonas and a method for selecting a bacterium belonging to the genus Pseudomonas through use of said selection medium.

A method for detecting, in a blood or tissue sample, dormant or cell wall deficient Mycobacterium species, said method including using a Ziehl Neelsen stain as hereinbefore defined, which includes treating the sample with carbol fuchsin, followed by treating the sample with a decolouriser and with a counter stain.

A testing method is described for use in undertaking a test upon a sample (10) for the presence of a specific microbiological material, the method comprising the steps of combining the sample (10) with a viability-preserving medium, optionally with an enrichment medium (12) selected to promote growth and/or reproduction of the specific microbiological material within the sample (10) and incubating the sample (10) and enrichment material (12), undertaking a separation process to separate the live specific microbiological material from the sample (10), and testing the separated material for the presence of the specific microbiological material. An apparatus for use in the method is also described.

The invention relates to the preparation of biological cells for the mass spectrometric analysis of cellular properties such as taxonomic classification, antibiotic resistances, response to drugs or other active substances, and others. The cells can be prokaryotic or eukaryotic microorganisms which have particularly been cultivated directly on a mass spectrometric sample support, or eukaryotic cells from tissues or cell cultures. The invention proposes that the cells are not disrupted by adding matrix solution for a subsequent ionization by matrix-assisted laser desorption (MALDI), but that they are disrupted in a separate treatment step using acids and/or solvents on the sample support itself. Surprisingly, the cell proteins released then adhere to the sample support so that they can be carefully washed with buffer solution to remove salts and other soluble impurities which can stem from earlier treatment steps, for example from nutrient solution.

A method for distinguishing among a first group of microorganisms belonging to a first taxon of Gram negative bacteria, the first group of bacteria exhibiting a mechanism of resistance to a treatment; a second group of microorganisms belonging to a second taxon of Gram negative bacteria, the second taxon of bacteria being different than said first taxon, and exhibiting a mechanism of resistance to a treatment identical to the mechanism of the first group; and a third group of Gram negative bacteria that is not resistant to the treatment.

The present disclosure provides rapid, convenient and sensitive means to detect low abundance target Gram negative bacteria such as Salmonella contamination from a range of samples, including food samples and environmental samples, through a single-step culture method and downstream sensitive detection.