A rapid DNA isolation procedure for the identification of Campylobacter jejuni by the polymerase chain reaction.
We have developed an efficient process for rapidly isolating campylobacter DNA using mechanical disruption combined with the guanidine-based reagent DNAzol. Template DNA was isolated by this method from cultures of Campylobacter jejuni resistant to lysis by boiling or enzymes and identified following polymerase chain reaction (PCR) amplification using primers specific for the hippuricase gene. Direct detection of campylobacters in poultry-processing samples by PCR is demonstrated in chicken carcass rinses spiked with lysis-resistant C. jejuni. Our results indicate that this method of DNA isolation may be ideal for direct PCR detection of pathogenic bacteria in complex samples of widely varied origin, especially when the target organisms are difficult to lyse by other means.
Englen
M D
MD
Poultry Microbiological Safety Research Unit, Richard B. Russell Agricultural Research Center, USDA Agricultural Research Service, Athens, GA 30604-5677, USA. menglen@saa.ars.usda.gov
Kelley
L C
LC
D000818:Animals; D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D005516:Food Microbiology; D008462:Meat-Packing Industry; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry / Q000382:microbiology*
D004269:DNA, Bacterial