Trypanosoma evansi: A comparison of PCR and parasitological diagnostic tests in experimentally infected mice.
Abstract: Trypanosoma evansi is the causative agent of equine trypanosomosis, disease that affects horse's productivity and health. Parasitological and molecular methods are mostly used to detect the infection. The aim of this work was evaluate PCR sensitivity to detect T. evansi using the primers 21/22-mer, ITS1, ESAG 6/7 and TBR 1/2 designed from repetitive (multicopies) genomic sequences. The results were compare with two parasitological tests in mice, micro-haematocrite centrifugation technique and direct microscopic examination. The results shows (a) that the minimum amount of DNA from blood of highly parasitaemic mice that was detectable by PCR was 0.001 ng, using the ESAG6/7 and TBR1/2 primer, (b) using TBR1/2 primer for parasites purified could detect 0.000001 ng and (c) in the prepatent period PCR detect the presence of parasites earlier than parasitological techniques. Nevertheless, the percentage of detection for PCR varies depending on primer employed with 60% and 66% for ITS1 and 21/22-mer, and 80% for ESAG6/7 and TBR1/2. Consequently, TBR1/2 and ESAG6/7 were the best primers to monitor T. evansi infections in mice. For epidemiological application, such comparative evaluation should be made for detection of T. evansi in livestock such as horses.
Publication Date: 2008-09-26 PubMed ID: 18848544DOI: 10.1016/j.exppara.2008.09.013Google Scholar: Lookup
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- Comparative Study
- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article compares the effectiveness of Polymerase Chain Reaction (PCR) and parasitological diagnostic tests in identifying Trypanosoma evansi infections in mice, with PCR proving more successful and the potential implications for horse health treatments in the future.
Understanding the Context
- Trypanosoma evansi is the parasite responsible for equine trypanosomosis, a disease that negatively impacts the health and productivity of horses.
- Currently, parasitological and molecular techniques are used most frequently to diagnose the infection.
- The main objective of this research was to gauge the sensitivity of PCR in detecting T. evansi in comparison to parasitological tests.
Methods and Materials Used
- The primers 21/22-mer, ITS1, ESAG 6/7 and TBR 1/2 were crafted from repetitive (multi-copies) genomic sequences to optimize PCR for detecting T. evansi.
- PCR results were juxtaposed with two parasitological tests conducted on mice: the micro-haematocrite centrifugation technique and direct microscopic examination.
Results of the Research
- The study determined that the minimum amount of DNA from the blood of highly parasitaemic mice detectable by PCR was 0.001 ng, achieved when using the ESAG6/7 and TBR1/2 primer.
- Furthermore, the usage of the TBR1/2 primer for purified parasites could detect 0.000001 ng.
- In the prepatent period, PCR detected the presence of parasites earlier than the parasitological techniques.
- However, the rate of detection for PCR differed based on the primer used, with ITS1 and 21/22-mer scoring 60% and 66% respectively, and ESAG6/7 and TBR1/2 scoring 80%.
Conclusions and Implications
- The research hence concluded that TBR1/2 and ESAG6/7 were the most effective primers to monitor T. evansi infections in mice.
- This indicates that PCR could potentially be a more sensitive method in detecting T. evansi, compared to traditional parasitological methods, which could have implications for equine health treatments.
- However, for these results to be applicable in a wider epidemiological context, similar comparative evaluations should be conducted with livestock such as horses.
Cite This Article
APA
Fernández D, González-Baradat B, Eleizalde M, González-Marcano E, Perrone T, Mendoza M.
(2008).
Trypanosoma evansi: A comparison of PCR and parasitological diagnostic tests in experimentally infected mice.
Exp Parasitol, 121(1), 1-7.
https://doi.org/10.1016/j.exppara.2008.09.013 Publication
Researcher Affiliations
- Universidad Nacional Experimental Simón Rodríguez, Instituto de Estudios Científicos y Tecnológicos, Centro de Estudios Biomédicos y Veterinarios, Apartado Postal 47925, Caracas 1041A, Venezuela.
MeSH Terms
- Animals
- Cattle
- Centrifugation
- DNA Primers / chemistry
- DNA, Protozoan / analysis
- DNA, Protozoan / blood
- Disease Models, Animal
- Equidae
- Female
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses
- Mice
- Parasitemia / diagnosis
- Parasitemia / parasitology
- Parasitemia / veterinary
- Polymerase Chain Reaction / standards
- Polymerase Chain Reaction / veterinary
- Rodentia
- Sensitivity and Specificity
- Trypanosoma / genetics
- Trypanosoma / isolation & purification
- Trypanosomiasis / diagnosis
- Trypanosomiasis / parasitology
- Trypanosomiasis / veterinary
Citations
This article has been cited 13 times.- Behour TS, Abd El Fattah EM. Genotyping of Trypanosoma brucei evansi in Egyptian camels: detection of a different non-RoTat 1.2 Trypanosoma brucei evansi in Egyptian camels.. Trop Anim Health Prod 2023 Jul 28;55(4):279.
- Javanshir A, Tavassoli M, Esmaeilnejad B. Morphological, serological, molecular detection, and phylogenetic analysis of Trypanosoma evansi in horses of different regions in Iran.. Parasitol Res 2023 Aug;122(8):1873-1881.
- Suprihati E, Suwanti LT, Yudhana A, Kusumaningrum AI. Comparison of ITS-1 and TBR-1/2 primer sensitivity for the detection of Trypanosoma evansi local isolates in experimental rats using a polymerase chain reaction.. Vet World 2022 Jul;15(7):1772-1778.
- Desquesnes M, Gonzatti M, Sazmand A, Thévenon S, Bossard G, Boulangé A, Gimonneau G, Truc P, Herder S, Ravel S, Sereno D, Jamonneau V, Jittapalapong S, Jacquiet P, Solano P, Berthier D. A review on the diagnosis of animal trypanosomoses.. Parasit Vectors 2022 Feb 19;15(1):64.
- Habeeb IF, Chechet GD, Kwaga JKP. Molecular identification and prevalence of trypanosomes in cattle distributed within the Jebba axis of the River Niger, Kwara state, Nigeria.. Parasit Vectors 2021 Oct 29;14(1):560.
- Setiawan A, Nurcahyo W, Priyowidodo D, Budiati RT, Susanti DSR. Genetic and parasitological identification of Trypanosoma evansi infecting cattle in South Sulawesi, Indonesia.. Vet World 2021 Jan;14(1):113-119.
- Behour TS, Aboelhadid SM, Mousa WM, Amin AS, El-Ashram SA. Molecular diagnosis of acute and chronic infection of Trypanosoma evansi in experimental male and female mice.. Onderstepoort J Vet Res 2019 Aug 26;86(1):e1-e10.
- Nassef NAE, El-Melegy MA, Beshay EV, Al-Sharaky DR, Al-Attar TM. Trypanocidal Effects of Cisplatin alone and in Combination with Nigella sativa Oil on Experimentally Infected Mice with Trypanosoma evansi.. Iran J Parasitol 2018 Jan-Mar;13(1):89-99.
- Ramírez-Iglesias JR, Eleizalde MC, Reyna-Bello A, Mendoza M. Molecular diagnosis of cattle trypanosomes in Venezuela: evidences of Trypanosoma evansi and Trypanosoma vivax infections.. J Parasit Dis 2017 Jun;41(2):450-458.
- Ramírez-Iglesias JR, Eleizalde MC, Gómez-Piñeres E, Mendoza M. Trypanosoma evansi: A clinical, parasitological and immunological evaluation of trypanosomosis using a chronic rabbit model.. Open Vet J 2012;2(1):78-82.
- Ybañez AP, Ybañez RH, Claveria FG, Cruz-Flores MJ, Xuenan X, Yokoyama N, Inokuma H. High genetic diversity of Anaplasma marginale detected from Philippine cattle.. J Vet Med Sci 2014 Jul;76(7):1009-14.
- Biryomumaisho S, Rwakishaya EK, Melville SE, Cailleau A, Lubega GW. Livestock trypanosomosis in Uganda: parasite heterogeneity and anaemia status of naturally infected cattle, goats and pigs.. Parasitol Res 2013 Apr;112(4):1443-50.
- Habila N, Agbaji AS, Ladan Z, Bello IA, Haruna E, Dakare MA, Atolagbe TO. Evaluation of In Vitro Activity of Essential Oils against Trypanosoma brucei brucei and Trypanosoma evansi.. J Parasitol Res 2010;2010.
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