title: Validation of the ANSR(®) Listeria monocytogenes Method for Detection of Listeria monocytogenes in Selected Food and Environmental Samples.
abstract: Work was conducted to validate performance of the ANSR(®) for Listeria monocytogenes method in selected food and environmental matrixes. This DNA-based assay involves amplification of nucleic acid via an isothermal reaction based on nicking enzyme amplification technology. Following single-step sample enrichment for 16-24 h for most matrixes, the assay is completed in 40 min using only simple instrumentation. When 50 distinct strains of L. monocytogenes were tested for inclusivity, 48 produced positive results, the exceptions being two strains confirmed by PCR to lack the assay target gene. Forty-seven nontarget strains (30 species), including multiple non-monocytogenes Listeria species as well as non-Listeria, Gram-positive bacteria, were tested, and all generated negative ANSR assay results. Performance of the ANSR method was compared with that of the U.S. Department of Agriculture, Food Safety and Inspection Service Microbiology Laboratory Guidebook reference culture procedure for detection of L. monocytogenes in hot dogs, pasteurized liquid egg, and sponge samples taken from an inoculated stainless steel surface. In addition, ANSR performance was measured against the U.S. Food and Drug Administration Bacteriological Analytical Manual reference method for detection of L. monocytogenes in Mexican-style cheese, cantaloupe, sprout irrigation water, and guacamole. With the single exception of pasteurized liquid egg at 16 h, ANSR method performance as quantified by the number of positives obtained was not statistically different from that of the reference methods. Robustness trials demonstrated that deliberate introduction of small deviations to the normal assay parameters did not affect ANSR method performance. Results of accelerated stability testing conducted using two manufactured lots of reagents predicts stability at the specified storage temperature of 4°C of more than 1 year.
authors.affiliation: Neogen Corp., 620 Lesher Pl, Lansing, MI 48912, USA.
authors.forename: Oscar
authors.initials: O
authors.lastname: Caballero
authors.forename: Susan
authors.initials: S
authors.lastname: Alles
authors.forename: Quynh-Nhi
authors.initials: QN
authors.lastname: Le
authors.forename: R Lucas
authors.initials: RL
authors.lastname: Gray
authors.forename: Edan
authors.initials: E
authors.lastname: Hosking
authors.forename: Lisa
authors.initials: L
authors.lastname: Pinkava
authors.forename: Paul
authors.initials: P
authors.lastname: Norton
authors.forename: Jerry
authors.initials: J
authors.lastname: Tolan
authors.forename: Mark
authors.initials: M
authors.lastname: Mozola
authors.forename: Jennifer
authors.initials: J
authors.lastname: Rice
authors.forename: Yi
authors.initials: Y
authors.lastname: Chen
authors.forename: Elliot
authors.initials: E
authors.lastname: Ryser
authors.forename: Joseph
authors.initials: J
authors.lastname: Odumeru
mesh_terms: D001431:Bacteriological Techniques; D004783:Environmental Microbiology; D005504:Food Analysis; D005516:Food Microbiology; D008089:Listeria monocytogenes / Q000235:genetics / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic; D014481:United States
chemical_list: D011933:Reagent Kits, Diagnostic