This technique, Magnetic Immuno PCR Assay (MIPA), is developed for the detection of Salmonella and other pathogens. The assay utilizes magnetic particles (MagneZoom) coated with monoclonal antibodies against Salmonella to extract these bacteria from a sample. Bacteria thus isolated are lysed, and the supernatant containining bacterial DNA, is subjected to the polymerase chain reaction (PCR) using primers from the Salmonella typhimurium origin of DNA replication to amplify a 163 bp region. The specificity of the primer set was tested in the PCR; amplification occurred with all 25Salmonella strains tested but not with other species of Enterobacteriaceae. A sensitivity of 100 cfu of Salmonella typhimurium was achieved for the MIPA by visualization of the amplified products by ethidium bromide stained agarose gel following electrophoresis. Further sensitivity can be increased by real tyime PCR or Southern blotting. The presence of Escherichia coli did not interfere with these detection levels. The MIPA specifically detected 100 cfu of Salmonella within 4-5 h time and may be potentially useful for rapid detection of Salmonella in clinical and food specimens.
Catalog No. | : | 10190002-1 | Size | : | 1 Kit |