Molecular detection of Clostridium botulinum type E neurotoxin gene in smoked fish by polymerase chain reaction and capillary electrophoresis.
The polymerase chain reaction (PCR), a rapid, sensitive technique for amplifying target DNA sequences of pathogenic microorganisms, 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.
Sciacchitano
C J
CJ
U.S. Food and Drug Administration, Northeast Regional Laboratory, Brooklyn, NY 11232, USA.
Hirshfield
I N
IN
D000818:Animals; D001483:Base Sequence; D001905:Botulinum Toxins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D003014:Clostridium botulinum / Q000235:genetics* / Q000378:metabolism; D003530:Cyprinidae; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D019075:Electrophoresis, Capillary; D005396:Fish Products / Q000382:microbiology*; D005506:Food Contamination; D005519:Food Preservation; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009498:Neurotoxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012474:Salmon; D012680:Sensitivity and Specificity; D014783:Viscosity
D004269:DNA, Bacterial; D009498:Neurotoxins; D001905:Botulinum Toxins