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DNA detection of Trypanosoma evansi : diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP)

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Abstract
Tiypanosoma evansi (T. evansi) is the most widely spread pathogenic trypanosome in the world. The control of trypanosomiasis depends on accurate diagnosis and effective treatment. Focusing on the presence of T. evansi in Asia, we developed a detection assay based on tracing phosphate ions (Pi) generated during LAMP targeting the variant surface glycoprotein (VSG) gene of Rode Trypanozoon antigenic type 1.2 (RoTat 1.2 VSG). The diagnostic potential as well as the use of the assay as a test-of-cure method after berenil treatment, was assessed in mice at different time points of infection. In addition, 67 buffalo blood collected from Tongling county, Anhui province, as well as 42 cattle sera from the Shanghai area, were used to evaluate the diagnostic validity of the test. The detection limit of the novel LAMP assay was determined to be as low as 1 fg of T. evansi DNA, while the reaction time for the test was only 30 min. Hence it outperforms both microscopy and PCR. In the test-of-cure assessment, successful berenil mediated cure could be confirmed within 48 h after treatment. This offers a tremendous advantage over conventional antibody-based diagnostic tools in which successful cure only can be confirmed after months. In the cattle and buffalo screening, the LAMP was able to detect a false-negative determined sample, wrongly classified in a conventional microscopy and PCR screening. Finally, no cross-reactivity was observed with other zoonotic parasites, such as T. evansi type B, T. congolense, T. brucei, Schistosoma japonicum, Plasmodium falciparum, Leishmania donovani, Toxoplasma gondii and Angiostrongylus cantonensis. We conclude that the novel LAMP assay is sensitive, specific and convenient for field use, particularly in areas where infection incidence has become extremely low. The LAMP assay could be used as a tool for trypanosomiasis control and elimination strategies in areas where T. evansi Type A infections are causing a threat to livestock fanning.
Keywords
Trypanosoma evansi, LAMP, Variant surface glycoprotein (VSG) gene, SENSITIVE DETECTION, WATER SAMPLES, DUPLEX PCR, INFECTIONS

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Citation

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MLA
Tong, Qunbo, Rui Chen, Qingming Kong, et al. “DNA Detection of Trypanosoma Evansi : Diagnostic Validity of a New Assay Based on Loop-mediated Isothermal Amplification (LAMP).” VETERINARY PARASITOLOGY 250 (2018): 1–6. Print.
APA
Tong, Q., Chen, R., Kong, Q., Goossens, J., Radwanska, M., Lou, D., Ding, J., et al. (2018). DNA detection of Trypanosoma evansi : diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP). VETERINARY PARASITOLOGY, 250, 1–6.
Chicago author-date
Tong, Qunbo, Rui Chen, Qingming Kong, Julie Goossens, Magdalena Radwanska, Di Lou, Jianzu Ding, et al. 2018. “DNA Detection of Trypanosoma Evansi : Diagnostic Validity of a New Assay Based on Loop-mediated Isothermal Amplification (LAMP).” Veterinary Parasitology 250: 1–6.
Chicago author-date (all authors)
Tong, Qunbo, Rui Chen, Qingming Kong, Julie Goossens, Magdalena Radwanska, Di Lou, Jianzu Ding, Bin Zheng, Yixiu Fu, Tianping Wang, Stefan Magez, and Shaohong Lu. 2018. “DNA Detection of Trypanosoma Evansi : Diagnostic Validity of a New Assay Based on Loop-mediated Isothermal Amplification (LAMP).” Veterinary Parasitology 250: 1–6.
Vancouver
1.
Tong Q, Chen R, Kong Q, Goossens J, Radwanska M, Lou D, et al. DNA detection of Trypanosoma evansi : diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP). VETERINARY PARASITOLOGY. 2018;250:1–6.
IEEE
[1]
Q. Tong et al., “DNA detection of Trypanosoma evansi : diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP),” VETERINARY PARASITOLOGY, vol. 250, pp. 1–6, 2018.
@article{8572064,
  abstract     = {Tiypanosoma evansi (T. evansi) is the most widely spread pathogenic trypanosome in the world. The control of trypanosomiasis depends on accurate diagnosis and effective treatment. Focusing on the presence of T. evansi in Asia, we developed a detection assay based on tracing phosphate ions (Pi) generated during LAMP targeting the variant surface glycoprotein (VSG) gene of Rode Trypanozoon antigenic type 1.2 (RoTat 1.2 VSG). The diagnostic potential as well as the use of the assay as a test-of-cure method after berenil treatment, was assessed in mice at different time points of infection. In addition, 67 buffalo blood collected from Tongling county, Anhui province, as well as 42 cattle sera from the Shanghai area, were used to evaluate the diagnostic validity of the test. The detection limit of the novel LAMP assay was determined to be as low as 1 fg of T. evansi DNA, while the reaction time for the test was only 30 min. Hence it outperforms both microscopy and PCR. In the test-of-cure assessment, successful berenil mediated cure could be confirmed within 48 h after treatment. This offers a tremendous advantage over conventional antibody-based diagnostic tools in which successful cure only can be confirmed after months. In the cattle and buffalo screening, the LAMP was able to detect a false-negative determined sample, wrongly classified in a conventional microscopy and PCR screening. Finally, no cross-reactivity was observed with other zoonotic parasites, such as T. evansi type B, T. congolense, T. brucei, Schistosoma japonicum, Plasmodium falciparum, Leishmania donovani, Toxoplasma gondii and Angiostrongylus cantonensis. We conclude that the novel LAMP assay is sensitive, specific and convenient for field use, particularly in areas where infection incidence has become extremely low. The LAMP assay could be used as a tool for trypanosomiasis control and elimination strategies in areas where T. evansi Type A infections are causing a threat to livestock fanning.},
  author       = {Tong, Qunbo and Chen, Rui and Kong, Qingming and Goossens, Julie and Radwanska, Magdalena and Lou, Di and Ding, Jianzu and Zheng, Bin and Fu, Yixiu and Wang, Tianping and Magez, Stefan and Lu, Shaohong},
  issn         = {0304-4017},
  journal      = {VETERINARY PARASITOLOGY},
  keywords     = {Trypanosoma evansi,LAMP,Variant surface glycoprotein (VSG) gene,SENSITIVE DETECTION,WATER SAMPLES,DUPLEX PCR,INFECTIONS},
  language     = {eng},
  pages        = {1--6},
  title        = {DNA detection of Trypanosoma evansi : diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP)},
  url          = {http://dx.doi.org/10.1016/j.vetpar.2017.12.006},
  volume       = {250},
  year         = {2018},
}

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