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The standard procedure for Shigella spp. detection is based on isolation of Shigella by selective culture media followed by identification by biochemical tests and agglutination assays. This process may take 48–72 h or even longer to obtain results. Since shigellae are very fastidious organisms, appropriate... 顯示更多 The standard procedure for Shigella spp. detection is based on isolation of Shigella by selective culture media followed by identification by biochemical tests and agglutination assays. This process may take 48–72 h or even longer to obtain results. Since shigellae are very fastidious organisms, appropriate collection, rapid transport to the laboratory and rapid plating of the sample are important for isolation. Such conditions are often difficult to attain in developing countries. Thus, rapid, highly sensitive and specific techniques based on genetic characteristics have been developed. DNA hybridization and PCR are the best known of these techniques and are often used as tests for the detection and identification of pathogenic microorganisms. Several PCR protocols for detection of Shigella in faeces, food, and some in water ( Bej and Bej) have been published. These protocols use primers directed at sequences located on the invasion plasmid of Shigella spp. and EIEC, or on both the plasmid and the chromosome . The major obstacle to using PCR for the detection and identification of pathogenic organisms from clinical samples or environmental water samples is the presence of substances that are inhibitory to PCR . This obstacle can be minimized by direct chemical extraction of nucleic acids, immunomagnetic separation (IMS) of bacteria with antibodies , or buoyant density gradient centrifugation, but these procedures are laborious and expensive.
In this paper, we describe the highly sensitive and specific detection of virulent Shigella organisms and EIEC by seminested PCR combined with a short enrichment step. The sensitivity of the procedure was determined by using artificially seeded ground and surface water samples collected from different natural sources.
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