Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Apr-Jun;13(2):225-234.

Development and Evaluation of a Loop-mediated Isothermal Amplification Assay for Rapid Detection of Theileria annulata Targeting the Cytochrome B Gene

Melek Chaouch  1 , Moez Mhadhbi  2 , Sassi Limam  2 , Mohamed Aziz Darghouth  2 , Souha Benabderrazak  1
Affiliations

Development and Evaluation of a Loop-mediated Isothermal Amplification Assay for Rapid Detection of Theileria annulata Targeting the Cytochrome B Gene

Melek Chaouch et al. Iran J Parasitol. 2018 Apr-Jun.

Abstract

Background: Theileria annulata is an economically important cattle disease in North Africa that occurs in subtropical and tropical areas. Accurate and rapid, molecular diagnosis of tropical theileriosis is an important issue that allows early treatment and, prevents transmission. We developed and validated a Theileria annulata specific LAMP assay targeting the cytochrome b multicopy gene, in order to increase the DNA detection sensitivity.

Methods: The methodology was used to evaluate the occurrences of T. annulata in 88 field samples collected in Northern Tunisia during 2013-2014. The specificity and sensitivity of the LAMP assays were compared to conventional cytochrome b PCR and routine microscopy commonly used on naturally infected cattle blood samples.

Results: The PCR assay showed a sensitivity of 70% and specificity around 75%. Our LAMP assay showed a suitable sensitivity 78.7% and specificity 87.5%, with, however, positive (98.4%) and negative (29.1%) predictive values.

Conclusion: The LAMP assay is a simple and convenient diagnostic tool for tropical theileriosis. Moreover, LAMP does not require experienced staff and specialized equipment for sampling procedures and it is practical outside laboratories and can be used for field diagnosis.

Keywords: Cytochrome b; Diagnosis; LAMP; Theileria annulata; Tunisia.

PubMed Disclaimer

Conflict of interest statement

Conflicts of interest The authors declare that there is no conflict of interest.

Figures

Fig. 1:
Fig. 1:
LAMP (A) and PCR (B) studied by agarose gel electrophoresis. Genome DNAs from 10-fold serially diluted T. annulata schizont-infected lymphocytes were used as templates
Fig. 2:
Fig. 2:
Amplification of different clinical DNA samples by loop-mediated isothermal amplification (LAMP) assays targeting the cytochrome b gene. (1) Positive control T. annulata DNA, (2-4-5) positives clinical samples, (3-6-7) Negatives clinical samples extracted from cattle blood, (8) H2O Negative control
Fig. 3:
Fig. 3:
Flow-diagram of the results obtained from diagnostics techniques applied to clinical samples. (TP is True Positive, TN represents True Negative, FN is False Negative and FP is False Positive)
See this image and copyright information in PMC

References

    1. Robinson PM. Theileriosis annulata and its transmission-a review. Trop Anim Health Prod. 1982; 14(1):3–12. - PubMed
    1. Darghouth MA, Bouattour A, Kilan M. Tropical theileriosis in Tunisia: epidemiology and control. Parassitologia. 1999; 41 Suppl 1:33–6. - PubMed
    1. Gharbi M, Sassi L, Dorchies P, Darghouth MA. Infection of calves with Theileria annulata in Tunisia: Economic analysis and evaluation of the potential benefit of vaccination. Vet Parasitol. 2006; 137(3–4):231–41. - PubMed
    1. Jeong W, Kweon CH, Kim JM, Jang H, Paik SG. Serological investigation of Theileria sergenti using latex agglutination test in South Korea. J Parasitol. 2005; 91(1):164–9. - PubMed
    1. Burridge MJ, Kimber CD, Young AS. Use of the indirect fluorescent antibody technique in serologic studies of Theileria lawrencei infections in cattle. Am J Vet Res. 1973;34(7):897–900. - PubMed

LinkOut - more resources