Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for Diagnosis of Tropical Theileriosis
- 8 July 2008
- journal article
- Published by Hindawi Limited in Transboundary and Emerging Diseases
- Vol. 55 (5-6), 238-243
- https://doi.org/10.1111/j.1865-1682.2008.01033.x
Abstract
A loop‐mediated isothermal amplification (LAMP) assay was developed and evaluated for diagnosis of tropical theileriosis. A set of six primers was designed based on the unique gene of Theileria annulata (Theileria annulata strain Ankara hypothetical protein (GeneDB TA04795). The protocol for the reaction was setup and the specificity and sensitivity of the assay were established. The specificity experiment showed that LAMP primers amplified T. annulata DNA successfully, while no amplification was seen for Theileria parva, Theileria mutans, Theileria sergenti, Theileria sinensis, Babesia bovis as well as bovine genomic DNA and water control. When the sensitivity of LAMP assay was compared with that of conventional PCR a 10‐fold higher sensitivity was found, with a detection limit of 10 pg/μl of genomic DNA isolated from a T. annulata‐infected cell line. The LAMP product was confirmed by restriction digestion and staining with SYBR Green I. Furthermore, the LAMP assay was applied for the diagnosis of T. annulata in field samples and compared with reverse line blot (RLB), demonstrating that results of the LAMP assay corresponded to those of RLB. These results indicate that the LAMP assay is rapid and simple to run, cost‐effective, sensitive and specific and has potential usefulness for application in epidemiological studies on T. annulata infection of cattle.Keywords
This publication has 20 references indexed in Scilit:
- Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosisVeterinary Parasitology, 2007
- Epidemiology ofTheileria annulataInfection of Dairy Cattle in the Sudan Using Molecular TechniquesAnnals of the New York Academy of Sciences, 2006
- Lamp for tadpolesNature Methods, 2005
- Molecular Evidence of Infections withBabesia gibsoniParasites in Japan and Evaluation of the Diagnostic Potential of a Loop-Mediated Isothermal Amplification MethodJournal of Clinical Microbiology, 2004
- Rapid Diagnosis of Human Herpesvirus 6 Infection by a Novel DNA Amplification Method, Loop-Mediated Isothermal AmplificationJournal of Clinical Microbiology, 2004
- Loop-Mediated Isothermal Amplification for Detection of African TrypanosomesJournal of Clinical Microbiology, 2003
- Loop-Mediated Isothermal Amplification for Direct Detection of Mycobacterium tuberculosis Complex, M. avium , and M. intracellulare in Sputum SamplesJournal of Clinical Microbiology, 2003
- Loop-mediated isothermal amplification of DNANucleic Acids Research, 2000
- Theileriasis : a comprehensive reviewRevue Scientifique et Technique de l'OIE, 1989
- Theilerial Species of Domestic LivestockPublished by Springer Nature ,1981