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Home / Equine Research Bank / Vet Parasitol / Development of loop-mediated isothermal amplification (LAMP)...
Veterinary parasitology2006; 143(2); 155-160; doi: 10.1016/j.vetpar.2006.08.014

Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosis.

Abstract: Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid method whereby DNA is amplified with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specifically designed primers and a DNA polymerase with strand displacement activity. In this study, we used LAMP primer sets designed from EMA-1 and Bc 48 genes for detection of Theileria equi and Babesia caballi infections, respectively. These primer sets specifically amplified DNA of the respective parasites. Both primer sets amplified T. equi and B. caballi up to 10(-6) dilution of 10-fold serially diluted samples. Furthermore, DNA extracted from blood collected from a horse experimentally infected with T. equi was amplified by a T. equi LAMP primer set from days 2 to 35 post-infection, demonstrating the high sensitivity of these primers. Of 55 samples collected from China, 81.8% and 56.3% were positively detected by LAMP for T. equi and B. caballi infections, respectively. In contrast, 91.8% and 45.9% of the 37 samples collected from South Africa were LAMP positive for T. equi and B. caballi, respectively. These results suggest that LAMP could be a potential diagnostic tool for epidemiological studies of equine piroplasmosis.
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Publication Date: 2006-09-14 PubMed ID: 16973284DOI: 10.1016/j.vetpar.2006.08.014Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research developed a novel nucleic acid method, Loop-mediated Isothermal Amplification (LAMP), to diagnose equine piroplasmosis with high specificity, efficiency, and rapidity. It suggests that the LAMP method can be an effective diagnostic tool for epidemiological studies of equine piroplasmosis.

Methodology

  • The researchers employed Loop-mediated isothermal amplification (LAMP) primer sets designed from EMA-1 and Bc 48 genes for the detection of Theileria equi (T. equi) and Babesia caballi (B. caballi) infections. T. equi and B. caballi are parasites that cause equine piroplasmosis.
  • The primer sets used exhibit a high level of specificity which means that they target and amplify specifically the DNA of T. equi and B. caballi. No cross-amplification with non-target DNA was observed.
  • These primer sets were able to amplify T. equi and B. caballi DNA down to 10(-6) dilution of 10-fold serially diluted samples, demonstrating the high sensitivity of these primer sets.

Results and Analysis

  • A T. equi LAMP primer set was used to amplify DNA extracted from a horse experimentally infected with T. equi between 2 to 35 days post-infection. This implies that the LAMP method can detect infections effectively in their early stages.
  • Among the 55 samples collected from China, 81.8% and 56.3% were detected positive by LAMP for T. equi and B. caballi infections, respectively.
  • On the other hand, 91.8% and 45.9% of the 37 samples collected from South Africa were confirmed positive by LAMP for T. equi and B. caballi, respectively.

Conclusion and Recommendations

  • Overall, the results suggest that the LAMP method could be a potential diagnostic tool for epidemiological studies of equine piroplasmosis due to its high efficiency, specificity, and rapidity.
  • The researchers recommend further studies to validate the potential use of the LAMP method in diagnosing equine piroplasmosis in other regions globally, taking into account the geographical variation in the prevalence of T. equi and B. caballi parasitic infections.

Cite This Article

APA
Alhassan A, Thekisoe OM, Yokoyama N, Inoue N, Motloang MY, Mbati PA, Yin H, Katayama Y, Anzai T, Sugimoto C, Igarashi I. (2006). Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosis. Vet Parasitol, 143(2), 155-160. https://doi.org/10.1016/j.vetpar.2006.08.014

Publication

Veterinary parasitology
ISSN: 0304-4017
NlmUniqueID: 7602745
Country: Netherlands
Language: English
Volume: 143
Issue: 2
Pages: 155-160

Researcher Affiliations

Alhassan, Andy
  • National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Thekisoe, Oriel M M
    Yokoyama, Naoaki
      Inoue, Noboru
        Motloang, Makhosazana Y
          Mbati, Peter A
            Yin, Hong
              Katayama, Yoshinari
                Anzai, Toru
                  Sugimoto, Chihiro
                    Igarashi, Ikuo

                      MeSH Terms

                      • Animals
                      • Babesia / isolation & purification
                      • Babesiosis / diagnosis
                      • Babesiosis / veterinary
                      • DNA Primers / chemistry
                      • DNA, Protozoan / chemistry
                      • Diagnosis, Differential
                      • Horse Diseases / diagnosis
                      • Horses
                      • Nucleic Acid Amplification Techniques / methods
                      • Nucleic Acid Amplification Techniques / standards
                      • Nucleic Acid Amplification Techniques / veterinary
                      • Polymerase Chain Reaction / methods
                      • Polymerase Chain Reaction / standards
                      • Polymerase Chain Reaction / veterinary
                      • Sensitivity and Specificity
                      • Species Specificity
                      • Theileria / isolation & purification
                      • Theileriasis / diagnosis
                      • Tick-Borne Diseases / diagnosis
                      • Tick-Borne Diseases / veterinary

                      Citations

                      This article has been cited 30 times.
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