title: Rapid detection of Salmonella in pet food: design and evaluation of integrated methods based on real-time PCR detection.
abstract: Reducing the risk of Salmonella contamination in pet food is critical for both companion animals and humans, and its importance is reflected by the substantial increase in the demand for pathogen testing. Accurate and rapid detection of foodborne pathogens improves food safety,  protects the public health, and benefits food producers by assuring product quality while facilitating product release in a timely manner. Traditional culture-based methods for Salmonella screening are laborious and can take 5 to 7 days to obtain definitive results. In this study, we  developed two methods for the detection of low levels of Salmonella in pet food using real-time PCR: (i) detection of Salmonella in 25 g of dried pet food in less than 14 h with an automated magnetic bead-based nucleic acid extraction method and (ii) detection of Salmonella  in 375 g of composite dry pet food matrix in less than 24 h with a manual centrifugation-based nucleic acid preparation method. Both methods included a preclarification step using a novel protocol that removes food matrix-associated debris and PCR inhibitors and improves the sensitivity  of detection. Validation studies revealed no significant differences between the two real-time PCR methods and the standard U.S. Food and Drug Administration Bacteriological Analytical Manual (chapter 5) culture confirmation method.
authors.affiliation: Life Technologies, Foster City, CA 94404, USA. priya.balachandran@lifetech.com
authors.forename: Priya
authors.initials: P
authors.lastname: Balachandran
authors.forename: Maria
authors.initials: M
authors.lastname: Friberg
authors.forename: V
authors.initials: V
authors.lastname: Vanlandingham
authors.forename: K
authors.initials: K
authors.lastname: Kozak
authors.forename: Amanda
authors.initials: A
authors.lastname: Manolis
authors.forename: Maxim
authors.initials: M
authors.lastname: Brevnov
authors.forename: Erin
authors.initials: E
authors.lastname: Crowley
authors.forename: Patrick
authors.initials: P
authors.lastname: Bird
authors.forename: David
authors.initials: D
authors.lastname: Goins
authors.forename: Manohar R
authors.initials: MR
authors.lastname: Furtado
authors.forename: Olga V
authors.initials: OV
authors.lastname: Petrauskene
authors.forename: Robert S
authors.initials: RS
authors.lastname: Tebbs
authors.forename: Duane
authors.initials: D
authors.lastname: Charbonneau
mesh_terms: D000821:Animal Feed / Q000382:microbiology*; D000818:Animals; D015169:Colony Count, Microbial / Q000379:methods*; D003257:Consumer Product Safety; D004269:DNA, Bacterial / Q000032:analysis; D005504:Food Analysis / Q000295:instrumentation* / Q000379:methods; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D060888:Real-Time Polymerase Chain Reaction; D012475:Salmonella / Q000302:isolation & purification*; D013997:Time Factors
chemical_list: D004269:DNA, Bacterial