Peer-reviewed veterinary case report
Development of a loop-mediated isothermal amplification method for detection of Theileria lestoquardi.
- Journal:
- Parasitology research
- Year:
- 2012
- Authors:
- Salih, Diaeldin A et al.
- Affiliation:
- Division of Veterinary Infection Biology and Immunology · Germany
Plain-English summary
Researchers have created a new test called a loop-mediated isothermal amplification (LAMP) assay to help diagnose infections caused by a parasite called Theileria lestoquardi. This test was designed to specifically identify the DNA of T. lestoquardi without mistakenly detecting other similar parasites. When tested on samples from sheep in Sudan, the LAMP assay showed a high level of accuracy, correctly identifying 92.1% of positive cases and 87.5% of negative cases when compared to another test called PCR. This new method could be a useful alternative for diagnosing T. lestoquardi infections in the field.
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.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/21744022/