Production of a sensitive immunoassay for detection of ruminant proteins in rendered animal material heated to &gt;130°C

Ansfield, Michael J.

doi:10.1080/09540109409354854

Cited by 26 publications

(28 citation statements)

References 8 publications

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“…Nevertheless, up to now the use of such an approach in eld conditions has led to some problems with regard to the reliability of the results, as indicated by an intercomparison study involving the testing of MBM in feed (1,29). To improve the reproducibility and sensitivity of this approach, we have introduced a faster and simpler commercially available method for DNA extraction and optimized the PCR procedure by selecting appropriate target templates.…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, there is currently no reliable, sensitive, and speci c method to detect the presence of the BSE causative agent in feeds. Therefore, only indirect approaches testing for the presence of animal constituents in ruminant feeds have been proposed and applied (1,16,17,(19)(20)(21)(22)28). Currently, the method of cially recommended by the EU for the control of MBM in animal feeds is the microscopic method, which involves the identi cation of an animal species through analysis of the tissue constituents (mainly bone fragments) on the basis of their morphological characteristics (7).…”

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confidence: 99%

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A Competitive Polymerase Chain Reaction–Based Approach for the Identification and Semiquantification of Mitochondrial DNA in Differently Heat-Treated Bovine Meat and Bone Meal

Frezza

Favaro

Vaccari

et al. 2003

Journal of Food Protection

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The risk of bovine spongiform encephalopathy propagation was drastically reduced after the European Union (EU) Health Authorities adopted restrictions involving a ban on animal-derived proteins in the diet of farm animals. Currently, the EU's of cially recommended method for controlling meat and bone meal (MBM) in animal feed is the microscopic method, which involves the identi cation of bone fragments on the basis of their morphological characteristics. Recently, we demonstrated that a polymerase chain reaction (PCR)-based assay can be used for the detection of taxon-speci c DNA in MBM and animal feeds. To ensure the safe rendering of animal by-products, the EU Council requires that this material be treated at 1338C at 300 kPa for 20 min. Here we investigate the relationship between DNA degradation, PCR ampli cation, and MBM heat treatment. With a competitive PCR-based approach, we compare the ampli cation ef ciency of bovine mitochondrial DNA target sequences of different lengths in several heat-treated MBM samples. For our method, a synthetic competitive DNA is used as an internal control for both DNA extraction and PCR reaction. A correlation between an increase in treatment temperature and a reduction in the size of the target sequences suitable for ampli cation was observed, suggesting progressive DNA fragmentation due to the temperature. We show that short amplicons (147 bp) can be used to detect the presence of bovine mtDNA in MBM samples treated according to the current European regulations. The use of such a competitive approach to compare ampli cation ef ciency levels of targets of different lengths might represent a useful tool for the determination of both the amount of MBM in animal feeds and its proper heat treatment.There is evidence that bovine spongiform encephalopathy (BSE) was spread around the world through animal feed containing infected meat and bone meal (MBM) (3, 4). The effective preventive measures taken by the United Kingdom with the introduction of the so-called ''real feed ban'' in 1996 (a ban on animal proteins in farm animal feeds) have led to a rapid reduction in the appearance of new cases in that country (5). Based on such epidemiological evidence, national and international authorities (Of ce International des Epizooties, the World Health Organization, Codex Alimentarius, and the European Commission) have progressively implemented speci c legislation and guidelines regarding the appropriate heat treatment of MBMs (10), the ban of MBMs from ruminant feeds (6), and the removal of speci ed risk materials from slaughtered ruminants (8). Currently, there is a temporary ban on animal meal in feed for all animals intended for human consumption within the European Union (EU), allowing mem- ber states to improve the practical implementation of the European legislation to prevent the spread of BSE (9). Nevertheless, there is currently no reliable, sensitive, and speci c method to detect the presence of the BSE causative agent in feeds. Therefore, only indirect approaches testing...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

A Competitive Polymerase Chain Reaction–Based Approach for the Identification and Semiquantification of Mitochondrial DNA in Differently Heat-Treated Bovine Meat and Bone Meal

Frezza

Favaro

Vaccari

et al. 2003

Journal of Food Protection

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“…Ansfield (5) was the first to explore the possibility of raising antibodies against thermo-stable antigens that are assumed to withstand severe rendering processes and to use them to check animal species. This led to a patented and in-house validated (6,7), reputed to be applicable to ruminant and porcine proteins in compound animal feedstuffs.…”

Section: Immunochemical Methodsmentioning

confidence: 99%

“…van Raamsdonk (2) , V. Baeten (3) , I. Murray (4) , G. Berben (3) , G. Brambilla (5) & C. von Holst (1) (1) European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Food and Feed Unit, Retieseweg, 2440 Geel, Belgium Since the consumption of infected material by ruminants has been unquestionably recognised as the main BSE transmission pathway, legislation has been established throughout the world to avoid MBM entering the ruminant feed chain. Proteins derived from mammalians were prohibited in ruminant feed in the European Union (EU) in 1994 (24) and in the United States of America (USA) in 1997 (74).…”

mentioning

confidence: 99%

An overview of tests for animal tissues in feeds applied in response to public health concerns regarding bovine spongiform encephalopathy

Gizzi

Raamsdonk²,

Baeten³

et al. 2003

Rev. Sci. Tech. OIE

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“…They are based on the analysis of proteins, e.g. isoelectric focussing (Jemmi & Schlosser, 1991;Jemmi & Schlosser, 1993;Wintero, Thomsen, & Davies, 1990), immunoassays (Ansfield, 1994;Hofmann, 1996;Hofmann, 1997;Hsieh, Sheu, & Bridgman, 1998;Morales et al, 1994) or gel diffusion (Waiblinger, Weber, & Kleinert, 1998).…”

Section: Introductionmentioning

confidence: 99%

In-house validation of a method for detection of animal meals in ruminant feeds by microscopy

et al. 2006

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Production of a sensitive immunoassay for detection of ruminant proteins in rendered animal material heated to >130°C

Cited by 26 publications

References 8 publications

A Competitive Polymerase Chain Reaction–Based Approach for the Identification and Semiquantification of Mitochondrial DNA in Differently Heat-Treated Bovine Meat and Bone Meal

A Competitive Polymerase Chain Reaction–Based Approach for the Identification and Semiquantification of Mitochondrial DNA in Differently Heat-Treated Bovine Meat and Bone Meal

An overview of tests for animal tissues in feeds applied in response to public health concerns regarding bovine spongiform encephalopathy

In-house validation of a method for detection of animal meals in ruminant feeds by microscopy

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