Rapid Screening for Salmonella in Raw Pet Food by Loop-Mediated Isothermal Amplification

Abstract

ABSTRACT Raw pet food, composed of raw meat and vegetables, has increased in popularity in recent years. Multiple surveys and frequent recalls indicate that this commodity has a high risk of contamination with Salmonella and other foodborne pathogens. Improved screening methods are needed to meet the growing demand for testing. This matrix verification study aimed to apply a Salmonella loop-mediated isothermal amplification (LAMP) method, recently completed multilaboratory validation in dry dog food, in several raw pet food matrices, following the U.S. Food and Drug Administration (FDA)'s method validation guidelines. Five types of raw pet food, consisting of freeze-dried beef and chicken treats and frozen beef, pork, and turkey complete foods, were evaluated. For each matrix, two sets of ten 25-g test portions (seven inoculated with ≤30 cells of Salmonella Typhimurium and three uninoculated controls) were examined. One set was preenriched in buffered peptone water and the other one was preenriched in lactose broth, which was followed by LAMP screening using two isothermal master mixes (ISO-001 and ISO-004). All results were confirmed by culture as specified in the FDA Bacteriological Analytical Manual (BAM). The LAMP method accurately detected Salmonella in all inoculated test portions of the five raw pet food samples, regardless of the preenrichment broth used. Positive results could be obtained within 4 min of the LAMP run using the ISO-004 master mix. All uninoculated controls tested negative using LAMP or BAM. In addition, one turkey-based complete pet food sample was found to be already contaminated with three Salmonella serovars harboring multiple antimicrobial resistance genes. The Salmonella LAMP method offers a rapid, reliable, and robust tool for routine screening of Salmonella in raw pet food, which will help better ensure product safety and protect public health. HIGHLIGHTS