title: Detection of central nervous system tissue on meat and carcass-splitting band saw blade surfaces using modified fluorescent glial fibrillary acidic protein enzyme-linked immunosorbent assay sampling and extraction procedures.
abstract: This study was conducted to determine optimal buffer pH, extraction procedure, and temperature for detecting central nervous system (CNS) tissue on meat surfaces and on carcass-splitting band saw blades using swab sampling. Glial fibrillary acidic protein (GFAP) is restricted to CNS tissue and has been used as a marker for CNS tissue presence in meat products. Sample preparation, extraction procedure, and extraction temperature of glial fibrillary acidic protein fluorescent enzyme-linked immunosorbent assay (GFAP F-ELISA) were modified to detect CNS tissue on meat surfaces and on carcass-splitting band saw blades. Maximum GFAP recovery was observed with an extraction buffer pH of 7.4. Extracting samples at room temperature by vortexing for 30 s in 1 ml of extraction buffer (phosphate-buffered saline [pH 7.4] plus 0.05% sodium dodecyl sulfate) consistently provided detection of GFAP on meat surfaces contaminated with 500 microg of spinal cord suspension per 50 cm2 and on carcass-splitting band saw blades contaminated with 20 microg of spinal cord suspension per 50 cm2. Recovery of GFAP was not affected by storing samples overnight at 4 degrees C. The current studies demonstrate the effectiveness of modified sampling procedures and preparations, sample extraction buffer pH, and extraction temperatures. These modifications introduced to the original F-ELISA sampling protocol result in asensitive and repeatable assay for detection of CNS tissue on meat surfaces and on carcass-splitting band saw blades.
authors.affiliation: Department of Animal Sciences, College of Agricultural Sciences, Colorado State University, Fort Collins, Colorado 80523, USA.
authors.forename: M C S
authors.initials: MC
authors.lastname: Reddy
authors.forename: Kim L
authors.initials: KL
authors.lastname: Hossner
authors.forename: Keith E
authors.initials: KE
authors.lastname: Belk
authors.forename: John A
authors.initials: JA
authors.lastname: Scanga
authors.forename: Robert S
authors.initials: RS
authors.lastname: Yemm
authors.forename: John N
authors.initials: JN
authors.lastname: Sofos
authors.forename: Gary C
authors.initials: GC
authors.lastname: Smith
mesh_terms: D000818:Animals; D003257:Consumer Product Safety; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004866:Equipment Contamination; D005506:Food Contamination / Q000032:analysis*; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D008460:Meat / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D005904:Glial Fibrillary Acidic Protein