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Bovine cysticercosis-Development of a real-time PCR to enhance classification of suspect cysts identified at meat inspection

Cuttell, L. and Owen, H. and Lew-Tabor, A. E. and Traub, R. J. (2013) Bovine cysticercosis-Development of a real-time PCR to enhance classification of suspect cysts identified at meat inspection. Veterinary Parasitology, 194 (1). pp. 65-69. ISSN 0304-4017

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Article Link(s): http://dx.doi.org/10.1016/j.vetpar.2013.02.018

Abstract

Laboratory confirmation methods are important in bovine cysticerosis diagnosis as other pathologies can result in morphologically similar lesions resulting in false identifications. We developed a probe-based real-time PCR assay to identify Taenia saginata in suspect cysts encountered at meat inspection and compared its use with the traditional method of identification, histology, as well as a published nested PCR. The assay simultaneously detects T. saginata DNA and a bovine internal control using the cytochrome c oxidase subunit 1 gene of each species and shows specificity against parasites causing lesions morphologically similar to those of T. saginata. The assay was sufficiently sensitive to detect 1 fg (Ct 35.09 +/- 0.95) of target DNA using serially-diluted plasmid DNA in reactions spiked with bovine DNA as well as in all viable and caseated positive control cysts. A loss in PCR sensitivity was observed with increasing cyst degeneration as seen in other molecular methods. In comparison to histology, the assay offered greater sensitivity and accuracy with 10/19 (53%) T. saginata positives detected by real-time PCR and none by histology. When the results were compared with the reference PCR, the assay was less sensitive but offered advantages of faster turnaround times and reduced contamination risk. Estimates of the assay's repeatability and reproducibility showed the assay is highly reliable with reliability coefficients greater than 0.94. Crown Copyright (C) 2013 Published by Elsevier B.V. All rights reserved.

Item Type:Article
Additional Information:Cuttell, Leigh Owen, Helen Lew-Tabor, Alicja E. Traub, Rebecca J. Meat and Livestock Australia (MLA) Project funded by Meat and Livestock Australia (MLA). We thank David Jordan (NSWDPI) and Baden Pearse (DAFF) for assistance and advice. Thanks to Jan Slapeta and Luzia Rast (USyd), John Ellis (USyd), Asma Latif (University of Punjab, Pakistan) and Tamsin Barnes (UQ) who supplied DNA or specimens of non-target parasites. Catherine Minchin (DAFF) performed the inter-laboratory assay reliability testing, Laercio Porto (UQ) assisted in statistical analysis and Jess Morgan (UQ) advised on assay design. We are grateful to the on-plant veterinarians for their help in forwarding suspect cysts identified at meat inspection. Elsevier science bv Amsterdam
Keywords:Bovine cysticercosis Taenia saginata Real-time PCR Meat inspection taenia-saginata cysticercosis diagnosis assay validation samples
Subjects:Animal culture > Cattle
Veterinary medicine > Veterinary pathology
Deposited On:28 Nov 2013 01:25
Last Modified:28 Nov 2013 01:25

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