title: Development of a loop-mediated isothermal amplification method for detection of Theileria lestoquardi.
abstract: A loop-mediated isothermal amplification (LAMP) assay was developed for the diagnosis of Theileria lestoquardi infection. The primers were designed based on the clone-5 sequence of T. lestoquardi. The specificity and sensitivity of the assay were established. Analysis of the specificity showed that the selected LAMP primers amplified the target sequence from T. lestoquardi DNA successfully, while no amplification was seen with DNA from Theileria annulata, Theileria ovis, Babesia ovis, Anaplasma ovis, or ovine genomic DNA. The specificity of the LAMP product was further confirmed by restriction digestion and sequencing. The sensitivity of the LAMP assay was analyzed in comparison to PCR resulting in a detection limit of 10 fg/μl of plasmid DNA containing the clone-5 sequence. The suitability for utilizing the LAMP assay in the field for the diagnosis of T. lestoquardi infection was tested on 100 field samples collected in Sudan and compared with results obtained by PCR. The relative specificity and sensitivity of the established LAMP assay was determined to be 92.1% and 87.5%, respectively, indicating that it may be regarded as an alternative molecular diagnostic tool to PCR which could be used for epidemiological surveys on T. lestoquardi infection.
authors: Division of Veterinary Infection Biology and Immunology, Research Center Borstel, Parkallee 22, 23845 Borstel, Germany.
authors: Diaeldin A
authors: DA
authors: Salih
authors: Awadia M
authors: AM
authors: Ali
authors: Zhijie
authors: Z
authors: Liu
authors: Mohammed A
authors: MA
authors: Bakheit
authors: Khalid M
authors: KM
authors: Taha
authors: Ahmed H
authors: AH
authors: El Imam
authors: Birgit
authors: B
authors: Kullmann
authors: Abdelrahim M
authors: AM
authors: El Hussein
authors: Jabbar S
authors: JS
authors: Ahmed
authors: Ulrike
authors: U
authors: Seitzer
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010274:Parasitology / Q000379:methods*; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis / Q000469:parasitology; D013397:Sudan; D016794:Theileria / Q000235:genetics / Q000302:isolation & purification*; D013801:Theileriasis / Q000175:diagnosis* / Q000469:parasitology; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D017931:DNA Primers