Novel Molecular Method for Detection of Bovine-Specific Central Nervous System Tissues as Bovine Spongiform Encephalopathy Risk Material in Meat and Meat.

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Novel Molecular Method for Detection of Bovine-Specific Central Nervous System Tissues as Bovine Spongiform Encephalopathy Risk Material in Meat and Meat Products Amir Abdulmawjood, Holger Schönenbrücher, Michael Bülte The Journal of Molecular Diagnostics Volume 7, Issue 3, Pages 368-374 (August 2005) DOI: 10.1016/S1525-1578(10)60566-1 Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Typical amplification products of bovine (lane 1), ovine (lane 2), and caprine (lane 3) GFAP cDNA with a size of 86 bp; negative RT-PCR of porcine GFAP cDNA (lane 4); positive PCR control by using genomic DNA with a size of 219 bp (lane 5); and negative control (lane 6); M, DNA molecular weight marker XIII 50-bp ladder (Roche). The Journal of Molecular Diagnostics 2005 7, 368-374DOI: (10.1016/S1525-1578(10)60566-1) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Relative standard curves show comparable amplification efficiencies of GFAP gene (target) PCR and 18S rRNA gene (housekeeping gene) real-time RT-PCR. The Δ slope was <0.1. The Journal of Molecular Diagnostics 2005 7, 368-374DOI: (10.1016/S1525-1578(10)60566-1) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Real-time GFAP RT-PCR amplification curves of undiluted and 10-, 100-, 1000-, 10,000-, and 100,000-fold dilutions of bovine cDNA. On the y axis, the absolute emission intensity is indicated; the x axis shows the number of PCR cycles. The calculated cDNA concentrations are shown in the table. The Journal of Molecular Diagnostics 2005 7, 368-374DOI: (10.1016/S1525-1578(10)60566-1) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 5 Detection of bovine GFAP mRNA in sausages with varying quantities of homogenized brain tissue added with (→) and without (→) heat treatment of the samples. The Journal of Molecular Diagnostics 2005 7, 368-374DOI: (10.1016/S1525-1578(10)60566-1) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 Relative quantitative value (Log10) of different organs calculated relative to the organ muscle, normalized to the housekeeping gene. The Journal of Molecular Diagnostics 2005 7, 368-374DOI: (10.1016/S1525-1578(10)60566-1) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions