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Evaluation of the Magnetic Immuno PCR assay for rapid detection ofSalmonella

  • M. N. Widjojoatmodjo
  • A. C. Fluit
  • R. Torensma
  • B. H. I. Keller
  • J. Verhoef
Article

Abstract

A new technique, the Magnetic Immuno PCR Assay (MIPA), has been developed for the detection ofSalmonella. The assay utilizes magnetic particles coated with monoclonal antibodies againstSalmonella to extract these bacteria from the sample. Trapped bacteria are lysed, and the supernatant, which contains bacterial DNA, is then subjected to the polymerase chain reaction (PCR) using primers from theSalmonella 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 19 other species ofEnterobacteriaceae tested. A sensitivity of 100 cfuSalmonella typhimurium was achieved for the MIPA by visualization of the amplified products by ethidiumbromide stained agarose gel electrophoresis. A tenfold higher sensitivity was obtained by Southern blotting of the amplified products. The presence of 107 cfuEscherichia coli did not interfere with these detection levels. The MIPA thus specifically detected 100 cfu ofSalmonella within 5 h and may be potentially useful for rapid detection ofSalmonella in clinical specimens and food.

Keywords

Polymerase Chain Reaction Monoclonal Antibody Agarose Ethidiumbromide Southern Blotting 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1991

Authors and Affiliations

  • M. N. Widjojoatmodjo
    • 1
    • 2
  • A. C. Fluit
    • 1
    • 2
  • R. Torensma
    • 1
    • 2
  • B. H. I. Keller
    • 3
  • J. Verhoef
    • 1
  1. 1.Eijkman Winkler Laboratory for Medical MicrobiologyUniversity Hospital UtrechtUtrechtThe Netherlands
  2. 2.U-gene Rescarch B.V.c/o University of UtrechtUtrechtThe Netherlands
  3. 3.Unilever Research LaboratoriumScction Microbiology and Hygienic ProcessingVlaardingenThe Netherlands

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