Detection of Bovine Central Nervous System Tissue as Bovine Spongiform Encephalopathy Risk Material by Real-Time Reverse Transcriptase-PCR in Raw and Cooked Beef Products

2008 ◽  
Vol 19 (4) ◽  
pp. 225-230 ◽  
Author(s):  
Xi-Ju Shi ◽  
Gui-Ping Ma ◽  
Bing-Ling Li ◽  
Jin-Liang Yang ◽  
Yan-Xin Li ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Reverse Transcriptase ◽  
Bovine Spongiform Encephalopathy ◽  
Real Time ◽  
Spongiform Encephalopathy ◽  
Central Nervous System Tissue ◽  
Reverse Transcriptase Pcr ◽  
Beef Products

Novel Approach for Detecting Prohibited Species-Specific Central Nervous System Tissue Contamination in Meat by One-Step Real-Time Reverse Transcriptase PCR

2009 ◽  
Vol 72 (5) ◽  
pp. 1063-1069 ◽  
Author(s):  
M. M. NAGARAJAN ◽  
D. LONGTIN ◽  
C. SIMARD
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Compliance Monitoring ◽  
Reverse Transcriptase Pcr ◽  
Qrt Pcr ◽  
Tissue Contamination ◽  
One Step ◽  
Species Specific

The dissemination of prohibited species-specific central nervous system (CNS) tissue contamination in meat must be tracked to mitigate human health risk associated with bovine spongiform encephalopathy. The efficiency of compliance monitoring and risk control measures taken by concerned regulatory authorities at meat production facilities to avoid such contamination depends on the ability to detect CNS tissue with a reliable and adequately sensitive quantitative method. A rapid and convenient one-step real-time quantitative reverse transcriptase PCR (qRT-PCR) assay was developed based on the absolute quantification of glial fibrillary acidic protein (GFAP) mRNA as a marker for CNS tissue contamination in meat. The GFAP RNA quantity corresponding to a percentage of CNS tissue in artificially spiked meat was determined using an appropriate in vitro transcribed target GFAP RNA as a calibration standard in the assay. The assay had a linear dynamic range of 102 to 109 copies of target RNA and was able to detect 0.01% CNS contamination in meat. Further evaluation consisted of an analysis of 272 random meat cuts from carcasses and 109 ground meat samples received from a federally inspected abattoir and two meat processing facilities, respectively, over a 5-month period. The analyzed samples were all negative for CNS tissue contamination at an arbitrarily set lower threshold of 0.025%. Overall, the newly developed one-step qRT-PCR may be useful as an objective quantitative compliance monitoring tool and for setting an acceptable low tolerance threshold for such contamination in meat.

Detection of Bovine Central Nervous System Tissues in Rendered Animal By-Products by One-Step Real-Time Reverse Transcription PCR Assay

2014 ◽  
Vol 77 (12) ◽  
pp. 2088-2097 ◽  
Author(s):  
OLGA ANDRIEVSKAIA ◽  
ERIN TANGORRA
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Bovine Spongiform Encephalopathy ◽  
Real Time ◽  
Binary Mixtures ◽  
Accurate Estimation ◽  
Spongiform Encephalopathy ◽  
Pcr Assay ◽  
Wide Range ◽  
By Products

Contamination of rendered animal byproducts with central nervous system tissues (CNST) from animals with bovine spongiform encephalopathy is considered one of the vehicles of disease transmission. Removal from the animal feed chain of CNST originated from cattle of a specified age category, species-labeling of rendered meat products, and testing of rendered products for bovine CNST are tasks associated with the epidemiological control of bovine spongiform encephalopathy. A single-step TaqMan real-time reverse transcriptase (RRT) PCR assay was developed and evaluated for specific detection of bovine glial fibrillary acidic protein (GFAP) mRNA, a biomarker of bovine CNST, in rendered animal by-products. An internal amplification control, mammalian β-actin mRNA, was coamplified in the duplex RRT-PCR assay to monitor amplification efficiency, normalize amplification signals, and avoid false-negative results. The functionality of the GFAP mRNA RRT-PCR was assessed through analysis of laboratory-generated binary mixtures of bovine central nervous system (CNS) and muscle tissues treated under various thermal settings imitating industrial conditions. The assay was able to detect as low as 0.05% (wt/wt) bovine brain tissue in binary mixtures heat treated at 110 to 130°C for 20 to 60 min. Further evaluation of the GFAP mRNA RRT-PCR assay involved samples of industrial rendered products of various species origin and composition obtained from commercial sources and rendering plants. Low amounts of bovine GFAP mRNA were detected in several bovine-rendered products, which was in agreement with declared species composition. An accurate estimation of CNS tissue content in industrial-rendered products was complicated due to a wide range of temperature and time settings in rendering protocols. Nevertheless, the GFAP mRNA RRT-PCR assay may be considered for bovine CNS tissue detection in rendered products in combination with other available tools (for example, animal age verification) in inspection programs.

Control Points To Reduce Movement of Central Nervous System Tissue during Beef Slaughter

2017 ◽  
Vol 80 (2) ◽  
pp. 355-360
Author(s):  
J. L. Aalhus ◽  
R. D. Thacker ◽  
I. L. Larsen ◽  
J. C. Roberts ◽  
M. A. Price ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Cold Water ◽  
Effective Means ◽  
Hot Water ◽  
Spongiform Encephalopathy ◽  
Cross Contamination ◽  
Central Nervous System Tissue ◽  
Before And After ◽  
Disease Variant

ABSTRACT Consumption of central nervous system tissue (CNST) from cattle with bovine spongiform encephalopathy (BSE) is thought to cause the human neurological disease, variant Creutzfeldt-Jacob disease. To identify points of cross-contamination of beef carcasses with CNST, 55 young beef cattle were slaughtered and processed through a federally inspected multispecies abattoir. The objectives of this study were to evaluate CNST spread following the placement of a plug in the penetration site of the skull after captive bolt stunning, to evaluate cross-contamination of carcasses before and after splitting, to compare the effects of hot water pasteurization (84°C for 10 s) versus cold water wash (10°C for 30 s) for reducing CNST on the carcass, and to examine other possible sources of cross-contamination in the abattoir. Results indicated that the use of a plastic plug reduced CNST contamination near the bolt penetration site. This study also confirmed that carcass splitting resulted in an increase in CNST contamination at various areas of the carcass. Hot water pasteurization appeared to be an effective means of removing CNST contamination from carcasses in most of the areas sampled.

Comparison of Analytical Methods for the Detection of Central Nervous System Tissue in Ground Beef

2008 ◽  
Vol 71 (7) ◽  
pp. 1505-1509
Author(s):  
DONG GYUN LIM ◽  
CHEORUN JO ◽  
MOOHA LEE
Keyword(s):  
Spinal Cord ◽  
Central Nervous System ◽  
Nervous System ◽  
Western Blotting ◽  
Ground Beef ◽  
Enzyme Linked Immunosorbent Assay ◽  
Spongiform Encephalopathy ◽  
Spinal Cord Tissue ◽  
Central Nervous System Tissue ◽  
By Products

Bovine spongiform encephalopathy is most likely transmitted by the consumption of central nervous system tissue of infected cows. The objective of this study was to compare the sensitivity or limits of detection of two central nervous system tissue detection assays (glial fibrillary acidic protein enzyme-linked immunosorbent assay [ELISA] and neuron-specific enolase Western blotting assay) in by-products and ground beef. By-products including brain, spinal cord, and ileum were collected from the slaughterhouse and used for analyses with ELISA and Western blotting assays. Beef samples were prepared by mixing ground beef with different amounts of spinal cord tissue (0, 0.03, 0.06, and 0.1%) and were analyzed using the two central nervous system tissue detection methods. Both analytical assays were applicable in detecting central nervous system tissue in ground beef. However, the ELISA method was considered superior because of its ease of use, high sensitivity, and rapidity as compared with the Western blot method.

scholarly journals Protease-resistant PrP deposition in brain and non-central nervous system tissues of a murine model of bovine spongiform encephalopathy

1996 ◽  
Vol 77 (8) ◽  
pp. 1941-1946 ◽  
Author(s):  
C. F. Farquhar ◽  
J. Dornan ◽  
R. C. Moore ◽  
R. A. Somerville ◽  
A. M. Tunstall ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Bovine Spongiform Encephalopathy ◽  
Murine Model ◽  
Spongiform Encephalopathy

Comparative Studies of a Real-Time PCR Method and Three Enzyme-Linked Immunosorbent Assays for the Detection of Central Nervous System Tissues in Meat Products†

2008 ◽  
Vol 71 (10) ◽  
pp. 2059-2066 ◽  
Author(s):  
HOLGER SCHÖNENBRÜCHER ◽  
KATRIN ANNETTE GÖBEL ◽  
AMIR ABDULMAWJOOD ◽  
JÜRGEN A. RICHT ◽  
MICHAEL BÜLTE
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Real Time ◽  
Real Time Pcr ◽  
Meat Products ◽  
Spongiform Encephalopathy ◽  
Pcr Assay ◽  
Heat Treated ◽  
Pcr Method ◽  
Minced Meat

The removal of certain central nervous system (CNS) tissues (part of the bovine spongiform encephalopathy risk material) from the food chain is one of the highest priority tasks associated with avoiding contamination of the human food chain with the agent of bovine spongiform encephalopathy. A recently developed real-time PCR assay and three commercially available enzyme-linked immunosorbent assays (ELISAs) for the detection of CNS tissues in minced meat and three types of heat-treated sausages were evaluated. Bovine brain was used for spiking of internal reference material, and its detectability was examined during storage times of 12 months (for frozen minced meat and liver sausage) and 24 months (for sausages treated with medium and high heat). The real-time PCR method and both ELISA kits detected 0.1% CNS tissue in frozen minced meat and 0.1 or 1% CNS tissue in heat-treated meat products. The detectability of the amplified mRNA target region with the PCR assay was similar to the detectability of antigen by the ELISAs. Because the real-time PCR method also can be used to distinguish cattle, ovine, and caprine CNS tissues from porcine CNS tissues, it seems to be suitable as a routine diagnostic test for the sensitive and specific detection of CNS tissues in meat and meat products.

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