Molecular detection of Clostridium botulinum type E neurotoxin gene in smoked fish by polymerase chain reaction and capillary electrophoresis
Sciacchitano, C.J.; Hirshfield, I.N.
Journal - Association of Official Analytical Chemists 79(4): 861-865
1996
ISSN/ISBN: 0004-5756 PMID: 8757444 Document Number: 456007
The polymerase chain reaction (PCR), a rapid, sensitive technique for amplifying target DNA sequences of pathogenic microorgansims, was used to amplify Clostridium botulinum type E neurotoxin gene fragments in smoked fish. Other botulinal neurotoxin-producing strains, nontoxigenic strains, and food-related microorganisms did not yield nonspecific amplification products with this PCR assay. PCR products were analyzed by capillary electrophoresis (CE) using a low-viscosity entangled polymer system. Resolution, sensitivity, and DNA sizing accuracy were improved, and analytical times were markedly shortened. The PCR/CE assay detected the C. botulinum type E neurotoxin gene in as few as 10 cells. The technique to other foods may also be a valuable tool for detecting foodborne pathogens.