Detection of Theileria annulata in blood samples of carrier cattle by PCR

d’Oliveira, Christine; Weide, Marjo van der; Habela, M.; Jacquiet, Philippe; Jongejan, Frans

doi:10.1128/jcm.33.10.2665-2669.1995

Cited by 208 publications

(84 citation statements)

References 19 publications

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“…Amplification was carried out by PCR targeting the small subunit (SSU) rRNA which is common to Theileria species [23]. All PCR reactions were performed in a volume of 25 µL with the primer pair, F (989) 5' AGT TTC TGA CCT ATC AG 3' and R (990) 5' TTG CCT TAA ACT TCC TTG 3' each at 3.2 µM.…”

Section: Pcr Amplification For Theileria Speciesmentioning

confidence: 99%

Haemoparasitic Infections in Cattle from a Trypanosoma brucei Rhodesiense Sleeping Sickness Endemic District of Eastern Uganda

et al. 2020

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We carried out a baseline survey of cattle in Kaberamaido district, in the context of controlling the domestic animal reservoir of Trypanosoma brucei rhodesiense human African trypanosomiasis (rHAT) towards elimination. Cattle blood was subjected to capillary tube centrifugation followed by measurement of the packed cell volume (PCV) and examination of the buffy coat area for motile trypanosomes. Trypanosomes were detected in 561 (21.4%) out of 2621 cattle screened by microscopy. These 561 in addition to 724 apparently trypanosome negative samples with low PCVs (≤25%) were transported to the laboratory and tested by PCR targeting the trypanosomal Internal Transcribed Spacer (ITS-1) as well as suspect Tick-Borne Diseases (TBDs) including Anaplasmamosis, Babesiosis, and Theileriosis. PCR for Anaplasma sp yielded the highest number of positive animals (45.2%), followed by Trypanosoma sp (44%), Theileria sp (42.4%) and Babesia (26.3%); multiple infections were a common occurrence. Interestingly, 373 (29%) of these cattle with low PCVs were negative by PCR, pointing to other possible causes of aneamia, such as helminthiasis. Among the trypanosome infections classified as T. brucei by ITS-PCR, 5.5% were positive by SRA PCR, and were, therefore, confirmed as T. b. rhodesiense. Efforts against HAT should therefore consider packages that address a range of conditions. This may enhance acceptability and participation of livestock keepers in programs to eliminate this important but neglected tropical disease. In addition, we demonstrated that cattle remain an eminent reservoir for T. b. rhodesiense in eastern Uganda, which must be addressed to sustain HAT elimination.

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Section: Pcr Amplification For Theileria Speciesmentioning

confidence: 99%

Haemoparasitic Infections in Cattle from a Trypanosoma brucei Rhodesiense Sleeping Sickness Endemic District of Eastern Uganda

et al. 2020

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“…The development of molecular biology has made accurate tools available for the detection of the parasite. A PCR assay for the diagnosis of T. annulata in the bovine host has been developed based on the Tams‐1 gene (d’Oliveira et al., 1995). Also, reverse line blotting (RLB) to detect and differentiate all known Theileria and Babesia species on the basis of differences in their 18S subunit rRNA gene sequences has been developed (Gubbels et al., 1999).…”

Section: Introductionmentioning

confidence: 99%

Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for Diagnosis of Tropical Theileriosis

Salih

Liu

Bakheit

et al. 2008

Transboundary and Emerging Diseases

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A loop-mediated isothermal amplification (LAMP) assay was developed and evaluated for diagnosis of tropical theileriosis. A set of six primers was designed based on the unique gene of Theileria annulata (Theileria annulata strain Ankara hypothetical protein (GeneDB TA04795). The protocol for the reaction was setup and the specificity and sensitivity of the assay were established. The specificity experiment showed that LAMP primers amplified T. annulata DNA successfully, while no amplification was seen for Theileria parva, Theileria mutans, Theileria sergenti, Theileria sinensis, Babesia bovis as well as bovine genomic DNA and water control. When the sensitivity of LAMP assay was compared with that of conventional PCR a 10-fold higher sensitivity was found, with a detection limit of 10 pg/microl of genomic DNA isolated from a T. annulata-infected cell line. The LAMP product was confirmed by restriction digestion and staining with SYBR Green I. Furthermore, the LAMP assay was applied for the diagnosis of T. annulata in field samples and compared with reverse line blot (RLB), demonstrating that results of the LAMP assay corresponded to those of RLB. These results indicate that the LAMP assay is rapid and simple to run, cost-effective, sensitive and specific and has potential usefulness for application in epidemiological studies on T. annulata infection of cattle.

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“…For the T. orientalis group, several sensitive PCR assays based on the gene encoding the major piroplasm surface protein have been developed (27,42). A similar approach using the Tams1 gene was followed for T. annulata (13). PCR detection of T. parva using repetitive DNA sequences or genespecific primers has also been developed (2).…”

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confidence: 99%

Simultaneous Detection of Bovine Theileria and Babesia Species by Reverse Line Blot Hybridization

et al. 1999

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A reverse line blot (RLB) assay was developed for the identification of cattle carrying different species ofTheileria and Babesia simultaneously. We included Theileria annulata, T. parva, T. mutans, T. taurotragi, and T. velifera in the assay, as well as parasites belonging to theT. sergenti-T. buffeli-T. orientalis group. TheBabesia species included were Babesia bovis,B. bigemina, and B. divergens. The assay employs one set of primers for specific amplification of the rRNA gene V4 hypervariable regions of all Theileria andBabesia species. PCR products obtained from blood samples were hybridized to a membrane onto which nine species-specific oligonucleotides were covalently linked. Cross-reactions were not observed between any of the tested species. No DNA sequences fromBos taurus or other hemoparasites (Trypanosomaspecies, Cowdria ruminantium, Anaplasma marginale, and Ehrlichia species) were amplified. The sensitivity of the assay was determined at 0.000001% parasitemia, enabling detection of the carrier state of most parasites. Mixed DNAs from five different parasites were correctly identified. Moreover, blood samples from cattle experimentally infected with two different parasites reacted only with the corresponding species-specific oligonucleotides. Finally, RLB was used to screen blood samples collected from carrier cattle in two regions of Spain. T. annulata, T. orientalis, and B. bigeminawere identified in these samples. In conclusion, the RLB is a versatile technique for simultaneous detection of all bovine tick-borne protozoan parasites. We recommend its use for integrated epidemiological monitoring of tick-borne disease, since RLB can also be used for screening ticks and can easily be expanded to include additional hemoparasite species.

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Detection of Theileria annulata in blood samples of carrier cattle by PCR

Cited by 208 publications

References 19 publications

Haemoparasitic Infections in Cattle from a Trypanosoma brucei Rhodesiense Sleeping Sickness Endemic District of Eastern Uganda

Haemoparasitic Infections in Cattle from a Trypanosoma brucei Rhodesiense Sleeping Sickness Endemic District of Eastern Uganda

Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for Diagnosis of Tropical Theileriosis

Simultaneous Detection of Bovine Theileria and Babesia Species by Reverse Line Blot Hybridization

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