Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for Diagnosis of Tropical Theileriosis
| Autor: | Diaeldin A. Salih, Jabbar S. Ahmed, Zhijie Liu, Mohammed A. Bakheit, Hermann Unger, A.M. El Hussein, Awadia M. Ali, G. Viljoen, Ulrike Seitzer |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Theileria parva
Molecular Sequence Data Loop-mediated isothermal amplification Polymerase Chain Reaction Sensitivity and Specificity Tropical theileriosis chemistry.chemical_compound parasitic diseases Animals DNA Primers Base Sequence General Veterinary General Immunology and Microbiology biology Babesia bovis General Medicine biology.organism_classification Theileria annulata Virology Molecular biology Theileriasis Blot genomic DNA chemistry SYBR Green I Cattle Restriction digest Nucleic Acid Amplification Techniques |
| Zdroj: | Transboundary and Emerging Diseases. 55:238-243 |
| ISSN: | 1865-1682 1865-1674 |
| DOI: | 10.1111/j.1865-1682.2008.01033.x |
| Popis: | Summary 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/ll 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|