Development of a loop-mediated isothermal amplification method for detection of Theileria lestoquardi

Division of Veterinary Infection Biology and Immunology, Research Center Borstel, Parkallee 22, 23845 Borstel, Germany.
Parasitology Research (Impact Factor: 2.1). 07/2011; 110(2):533-8. DOI: 10.1007/s00436-011-2518-x
Source: PubMed


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.

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Available from: Diaeldin A Salih,
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    • "In addition, the reaction can be carried out without the need of DNA extraction. The method has been successfully developed for the detection of several TBDs (Salih et al., 2008; Liu et al., 2008; Muller et al., 2010; Thekisoe et al., 2010;Salih et al., 2012). "

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    • "Furthermore, the extracted DNA was tested for natural T. lestoquardi infection using Loop-mediated isothermal amplification (LAMP) test. The LAMP primers and the procedure followed in reaction condition and the detection of the reaction products were essentially as described by Salih et al. (2012). "
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