DNA detection of Trypanosoma evansi: Diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP)
| Autor: | Ding Jianzu, Shaohong Lu, Julie Goossens, Magdalena Radwanska, Bin Zheng, Di Lou, Qunbo Tong, Tianping Wang, Yixiu Fu, Magez Stefan, Qingming Kong, Rui Chen |
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| Přispěvatelé: | Department of Bio-engineering Sciences, Cellular and Molecular Immunology, Faculty of Sciences and Bioengineering Sciences |
| Rok vydání: | 2018 |
| Předmět: |
Veterinary Medicine
0301 basic medicine China Trypanosoma 030231 tropical medicine 030106 microbiology Loop-mediated isothermal amplification Leishmania donovani Cattle Diseases Sensitivity and Specificity Mice 03 medical and health sciences 0302 clinical medicine Limit of Detection Trypanosomiasis parasitic diseases Journal Article medicine Animals General Veterinary biology Schistosoma japonicum Toxoplasma gondii Plasmodium falciparum General Medicine DNA Protozoan Trypanosoma evansi biology.organism_classification medicine.disease Virology Angiostrongylus cantonensis Cattle Parasitology Nucleic Acid Amplification Techniques Variant Surface Glycoproteins Trypanosoma |
| Zdroj: | Veterinary Parasitology. 250:1-6 |
| ISSN: | 0304-4017 |
| DOI: | 10.1016/j.vetpar.2017.12.006 |
| Popis: | Trypanosoma 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 30min. Hence it outperforms both microscopy and PCR. In the test-of-cure assessment, successful berenil mediated cure could be confirmed within 48h 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 farming. |
| Databáze: | OpenAIRE |
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