Abstract
The diagnosis of gastrointestinal nematode infections in small ruminants is central to studying the biology and epidemiology of these parasites and underpins their control. Traditional methods of diagnosis are inaccurate, time-consuming and laborious. Here, we describe a step-by-step protocol for the molecular-based diagnosis of infections by real-time PCR.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Perry BD, Randolph TF (1999) Improving the assessment of the economic impact of parasitic diseases and of their control in production animals. Vet Parasitol 84:145–168
Sackett D, Holmes P (2006) Assessing the economic cost of endemic disease on the profitability of Australian beef cattle and sheep producers. Meat and Livestock Australia, Report No. ISBN: 1741910021, Sydney, Australia
Hoste H, Torres-Acosta JF (2011) Non chemical control of helminths in ruminants: adapting solutions for changing worms in a changing world. Vet Parasitol 180:144–154
Wolstenholme AJ, Fairweather I, Prichard R, Samson-Himmelstjerna GV, Sangster NC (2004) Drug resistance in veterinary helminths. Trends Parasitol 20:469–476
Kaplan RM (2004) Drug resistance in nematodes of veterinary importance: a status report. Trends Parasitol 20:477–481
Gasser RB, Bott NJ, Chilton NB, Hunt P, Beveridge I (2008) Towards practical, DNA-based diagnostic methods for parasitic nematodes of livestock-bionomic and bio-technological implications. Biotechnol Adv 26:325–334
Roeber F, Jex AR, Gasser RB (2013) Next-generation molecular-diagnostic tools for gastrointestinal nematodes of livestock, with emphasis on small ruminants: a turning point? Adv Parasitol 83:267–333
Bott NJ, Campbell BE, Beveridge I, Chilton NB, Rees D, Hunt PW et al (2009) A combined microscopic-molecular method for the diagnosis of strongylid infections in sheep. Int J Parasitol 39:1277–1287
Roeber F, Jex AR, Campbell AJD, Campbell BE, Anderson GA, Gasser RB (2011) Evaluation and application of a molecular method to assess the composition of strongylid nematode populations in sheep with naturally acquired infections. Infect Genet Evol 11:849–854
Roeber F, Larsen JWA, Anderson N, Campbell AJD, Anderson GA, Gasser RB et al (2012) A molecular diagnostic tool to replace larval culture in conventional faecal egg count reduction testing in sheep. PLoS One 7:e37327
Gasser RB, Hu M, Chilton N, Campbell BE, Jex AR, Otranto D et al (2006) Single-strand conformation polymorphism (SSCP) for the analysis of genetic variation. Nat Protoc 1:3121–3128
Taylor MA, Coop RL, Wall RL (2007) The laboratory diagnosis of parasitism. Chapter 15. In: Taylor MA, Coop RL, Wall RL (eds) Veterinary parasitology, 3rd edn. Blackwell, Oxford, UK, p 798
Kemp B, Smith D (2005) Use of bleach to eliminate contaminating DNA from the surface of bones and teeth. Forensic Sci Int 154:53–61
Roeber F, Jex AR, Campbell AJ, Nielsen R, Anderson GA, Stanley KK et al (2012) Establishment of a robotic, high-throughput platform for the specific diagnosis of gastrointestinal nematode infections in sheep. Int J Parasitol 42:1151–1158
Acknowledgments
This research was supported through funds from the Australian Research Council (RBG and ARJ). Other support from Melbourne Water Corporation, the National Health and Medical Research Council, and the Alexander von Humboldt Foundation is gratefully acknowledged (RBG). FR was the recipient of scholarships from the University of Melbourne.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Roeber, F., Jex, A.R., Gasser, R.B. (2015). A Real-Time PCR Assay for the Diagnosis of Gastrointestinal Nematode Infections of Small Ruminants. In: Cunha, M., Inácio, J. (eds) Veterinary Infection Biology: Molecular Diagnostics and High-Throughput Strategies. Methods in Molecular Biology, vol 1247. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2004-4_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2004-4_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2003-7
Online ISBN: 978-1-4939-2004-4
eBook Packages: Springer Protocols