Pathogens culture methods

As well as overcoming many of the inherent problems associated with agriculture, plant tissue culture also offers a number of advantages over conventional animal cell culture methods currently being applied to produce biopharmaceutical proteins commercially [8], As plant culture media are relatively simple in composition and do not contain proteins, the cost of the process raw materials is reduced and protein recovery from the medium is easier and cheaper compared with animal cell culture. In addition, as most plant pathogens are unable to infect humans, the risk of pathogenic infections being transferred from the cell culture via the product is also substantially reduced. 

The FDA, USDA, and EPA have established methods and standards for detecting food- and water-borne pathogens these methods are available on the agencies websites [4—81. Many of the procedures for microbe identification rely on culturing the sample for subsequent identification by multiple tests such as colony characteristics, selective growth conditions, and biochemical assays for metabolites. Such traditional methods take upward of 24 h and remain the standard against which new... 

These growing and enrichment steps are relatively time-consuming, having a total assay time of up to 1 week in certain food pathogens [6]. Although classical culture methods can be sensitive, they are greatly restricted by the assay time. 

Laboratory tests are an Important adjunct in confirming the presence of suspected bioterror agents. Laboratory identification requires several approaches (Hen-chal, Teska, Ludwig, Shoemaker, Ezzell, 2001). One component is the use of culture methods. For the bacterial pathogens of interest [Bacillus anthracis, Yersinia pestis, and Frandsella tularensis), initial staining and microscopy results are available rapidly, and can assist... 

A wide range of pathogenic bacteria can be detected in water samples, usually by culture methods. Most commonly, quantitative tests are carried out that require several steps preenrichment (= resuscitation), selective enrichment, isolation by plate culture, identification by biochemical and/or serological tests, and eventually epidemiological typing. 

To ensure the safety of food products, representative samples must be inspected so that foodborne bacteria can be identified.15,18,19 Bacteria producing heat-stable enterotoxins, such as Staphylococcus aureus, may be identified by biochemical and serological techniques.20,21 Molecular methods are now widely used for the identification of many pathogenic foodborne bacteria,15,22,23 In addition bacteria used as starter cultures for cheese, yogurt, other fermented foods and beverages, and probiotic dietary supplements may be identified for quality assurance.22,24,25... 

The limitations of ELISA methods include the specificity of antibodies, the concentrations of primary antibody and antigen, and the type of reaction solution. Nonspecific binding of either of the antibodies to related antigens, unrelated proteins of other bacteria, or even the microtiter plate may lead to false positive reactions.49,52 57 Use of a monoclonal antibody may decrease crossreactivity with other antigens. For detection of low numbers of bacteria, as in drinking water, the sample may be filtered to concentrate the cells or cultured in a selective broth until it reaches the minimum detection limit for ELISA.49,58 Commercial test kits using dipsticks, immunoblots, and sandwich ELISA methods have been developed for the identification of pathogenic bacteria.58,59... 

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