PCR–RFLP using Ssu-rDNA amplification as an easy method for species-specific diagnosis of Trypanosoma species in cattle

Geysen, Dirk; Delespaux, V.; Geerts, S.

doi:10.1016/s0304-4017(02)00313-8

Cited by 110 publications

(90 citation statements)

References 14 publications

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“…The full length 18S ribosomal DNA sequence was analyzed using the forward primer 18STnF2 (5'-CAA CGA TGA CAC CCA TGA ATT GGG GA-3') and 18STnR3 (5'- TGC GCG ACC AAT AAT TGC AAT AC-3') as reverse primer [7]. For single PCR amplification, Kin1 reverse (5'-GCG TTC AAA GAT TGG GCA AT-3') and Kin2 forward (5'-CGC CCG AAA GTT CAC C-3') primers were used to anneal internal transcribed spacer of ribosomal genes (ITS) sequence, the conserved spacer between the 5.8S and 18S ribosomal DNA [3,17].…”

Section: Cryo-preservation and Long-term Continuous Cultivationmentioning

confidence: 99%

“…Owing to the lack of a reliable serological diagnostic method for this species and a typically low parasitemia, cultivation of blood samples is better method for detection. Furthermore, T. theileri can be distinguished from other trypanosomes by PCR-amplified spliced-leader transcript [8], internal transcribed spacer of ribosomal genes (ITS gene) [3], 18S ribosomal DNA [7] and a species specific primer set Tth625a,b [19]. However, the spliced-leader transcript is not an informative phylogenetic marker for long range evolutionary studies of the genus Trypanosoma [8], hence we amplified and sequenced the other three genes to identify and characterize T. theileri isolated from the cattle in Taiwan 1 (TWTth1) in the current study.…”

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confidence: 99%

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Isolation of Trypanosoma (Megatrypanum) theileri from Dairy Cattle in Taiwan

et al. 2010

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ABSTRACT. Although Trypanosoma (Megatrypanum) theileri, a blood parasite of bovid species, is spread widely throughout the world, it has never been reported in Taiwan. When an anti-coagulated blood sample from febrile dairy cattle was directly smeared, no parasite was observed. However, a highly distinctive morphological feature of trypanosome appeared in baby hamster kidney (BHK) cell culture inoculated with non-thrown blood buffy coat. The different stages and typical ultrastructures of trypanosome were observed in our isolate. The isolate was subsequently identified as T. theileri by species-specific PCR assay (Tth625), 18S rDNA sequencing alignment and internal transcribed spacer of ribosomal genes (ITS) as a marker for molecular phylogenetic analysis. The first T. theileri isolate in Taiwan (TWTth1) could be periodically passaged in BHK cell culture for more than one year and retained good re-cryopreservation viability. The BHK culture method would be excellent for diagnostic isolation and maintenance long-term development of this parasite.

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Section: Cryo-preservation and Long-term Continuous Cultivationmentioning

confidence: 99%

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confidence: 99%

Isolation of Trypanosoma (Megatrypanum) theileri from Dairy Cattle in Taiwan

et al. 2010

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“…PCR has been applied for the detection of many species of Babesia and Theileria and has been shown to have higher sensitivity and specificity compared with serological assays (Geysen et al, 2003, Buling et al, 2007, Jefferies et al, 2007and Sibeko et al, 2008.…”

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confidence: 99%

Sequence heterogeneity in the 18S rRNA gene within Theileria equi and Babesia caballi from horses in South Africa

Bhoora

Franssen

Oosthuizen

et al. 2009

Veterinary Parasitology

131

126

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“…The primers were adopted with minor modifications from Geysen et al (2003). All primers were purchased from InVitrogen.…”

Section: Pcr Screening Of Blood Samplesmentioning

confidence: 99%

“…Ribosomal DNA has previously been shown to be useful for trypanosome analysis (Geysen et al 2003;Desquesnes and Dávila 2002).…”

Section: Introductionmentioning

confidence: 99%

Trypanosoma spp. in Swedish game animals

2011

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Serum and blood samples from 36 game animals, shot during the hunting seasons 2007-2009, were collected and analyzed for the presence of Trypanosoma spp. by three methods; Isolation, PCR and serology. Only fisseped animals were included, four different ruminants and wild boar. Trypanosomes could be isolated from two of the animals, and eight had detectable parasite DNA. Seven animals had high titers of anti trypanosoma IgG antibodies. The two isolated strains, one from roe dear and one from European elk, were determined to T. theileri by partial DNA sequencing of the 18 S ribosomal gene. In the seven boars no Trypanosoma were detected, but 4 out of 7 strongly positive serological samples came from this group. This is the first study in Scandinavia on the presence of Trypanosoma in game animals. The results indicate that trypanosomiasis is frequently occurring among Swedish game animals.

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PCR–RFLP using Ssu-rDNA amplification as an easy method for species-specific diagnosis of Trypanosoma species in cattle

Cited by 110 publications

References 14 publications

Isolation of Trypanosoma (Megatrypanum) theileri from Dairy Cattle in Taiwan

Isolation of Trypanosoma (Megatrypanum) theileri from Dairy Cattle in Taiwan

Sequence heterogeneity in the 18S rRNA gene within Theileria equi and Babesia caballi from horses in South Africa

Trypanosoma spp. in Swedish game animals

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