Differentiation of two ovine Babesia based on the ribosomal DNA internal transcribed spacer (ITS) sequences

Niu, Qingli; Luo, Jianxun; Guan, Guiquan; Liu, Zhijie; Ma, Miling; Liu, Aihong; Gao, Jinliang; Ren, Qiaoyun; Li, Youquan; Jiaxiang, Qiu; Yin, Hong

doi:10.1016/j.exppara.2008.09.021

Cited by 45 publications

(28 citation statements)

References 15 publications

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“…We targeted the ITS regions of nuclear ribosomal RNA given the divergence in sequence variation among closely related Babesia species (Blaschitz et al, 2008; Bostrom et al, 2008; Liu et al, 2008; Niu et al, 2009). Quantitative data of these assays were compared with real-time PCR assays using identical primers and probes.…”

Section: Introductionmentioning

confidence: 99%

Development of droplet digital PCR for the detection of Babesia microti and Babesia duncani

Wilson

Glaser

Adams-Fish

et al. 2015

Experimental Parasitology

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Babesia spp. are obligate protozoan parasites of red blood cells. Transmission to humans occurs through bites from infected ticks or blood transfusion. Infections with B. microti account for the majority of the reported cases of human babesiosis in the USA. A lower incidence is caused by the more recently described species B. duncani. The current gold standard for detection of Babesia is microscopic examination of blood smears. Recent PCR-based assays, including real-time PCR, have been developed for B. microti. On the other hand, molecular assays that detect and distinguish between B. microti and B. duncani infections are lacking. Closely related species of Babesia can be differentiated due to sequence variation within the internal transcribed spacer (ITS) regions of nuclear ribosomal RNAs. In the present study, we targeted the ITS regions of B. microti and B. duncani to develop sensitive and species-specific droplet digital PCR (ddPCR) assays. The assays were shown to discriminate B. microti from B. duncani and resulted in limits of detection of ~10 gene copies. Moreover, ddPCR for these species were useful in DNA extracted from blood of experimentally infected hamsters, detecting infections of low parasitemia that were negative by microscopic examination. In summary, we have developed sensitive and specific quantitative ddPCR assays for the detection of B. microti and B. duncani in blood. Our methods could be used as sensitive approaches to monitor the progression of parasitemia in rodent models of infection as well as serve as suitable molecular tests in blood screening.

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Section: Introductionmentioning

confidence: 99%

Development of droplet digital PCR for the detection of Babesia microti and Babesia duncani

Wilson

Glaser

Adams-Fish

et al. 2015

Experimental Parasitology

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“…Unlike the rRNA genes which are highly conserved between closely related species, the spacer regions (ITS1 and ITS2) are subject to higher evolutionary rates and are therefore more variable in their lengths and nucleotide composition (Hillis and Dixon, 1991). These regions have therefore been used for the discrimination of closely related species and subspecies, and in the description of new species (Zahler et al, 1998;Holman et al, 2003;Lew et al, 2003;Aktas et al, 2007;de Rojas et al, 2007;Hilpertshauser et al, 2007;Saito-Ito et al, 2008;Niu et al, 2009;Bosman et al, 2010).…”

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confidence: 99%

Phylogeny of Theileria buffeli genotypes identified in the South African buffalo (Syncerus caffer) population

Chaisi

Collins

Oosthuizen

2014

Veterinary Parasitology

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Theileria buffeli/T. sergenti/T. orientalis is a group of benign and mildly pathogenic species of cattle and buffalo. In a previous study, we identified T .buffeli in blood samples originating from the African buffalo (Syncerus caffer) in the Hluhluwe-iMfolozi Game Park (HIP) and the Addo Elephant Game Park (AEGP) in South Africa. The aim of this study was to characterize the 18S rRNA gene and complete internal transcribed spacer (ITS1-5.8S-ITS2) region of T. buffeli samples, and to establish the phylogenetic position of this species based on these loci. The 18S rRNA gene and the complete ITS region were amplified from DNA extracted from blood samples originating from buffalo in these localities. The PCR products were cloned and the resulting recombinants sequenced. We identified novel T. buffeli 18S rRNA gene and ITS genotypes from buffalo in the AEGP, and novel T. sinensis 18S rRNA genotypes from buffalo in the HIP. Phylogenetic analyses indicated that the T. buffeli-like sequences were similar to T. buffeli sequences from cattle and buffalo in China and India, and the T. sinensis-like sequences were similar to T. sinensis 18S rRNA sequences of cattle and yak in China. There was extensive sequence variation between the novel T.1 buffeli genotypes of the African buffalo and previously described T. buffeli and T. sinensis genotypes. The presence of organisms with T. buffeli-like and T. sinensis-like genotypes in the African buffalo could be of significant importance, particularly to the cattle industry in South Africa as these animals might act as sources of infections to naïve cattle. This is the first report on the characterization of the full-length 18S rRNA gene and ITS region of T. buffeli and T. sinensis genotypes in South Africa. Our study provides invaluable information towards the classification of this complex group of benign and mildly pathogenic species.

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“…T. luwenshuni and T. uilenbergi shared the same shape, host, vector tick, pathogenicity, and distribution, but their phylogenetic relationships were distant in the 28S rRNA and D2-D3 segment trees. These results also provided evidence that T. luwenshuni and T. uilenbergi are 2 different and valid species [30,31]. …”

Section: Discussionmentioning

confidence: 77%

“…The 18S rRNA gene is used most commonly in studies of Theileria but it has a high degree of conservation and does not always discriminate among closely related species [30,31]. The 28S rRNA gene comprises tandem conserved and diverse regions.…”

Section: Discussionmentioning

confidence: 99%

Phylogenetic Analysis of Ruminant Theileria spp. from China Based on 28S Ribosomal RNA Gene

Gou

Guan

et al. 2013

Korean J Parasitol

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Species identification using DNA sequences is the basis for DNA taxonomy. In this study, we sequenced the ribosomal large-subunit RNA gene sequences (3,037-3,061 bp) in length of 13 Chinese Theileria stocks that were infective to cattle and sheep. The complete 28S rRNA gene is relatively difficult to amplify and its conserved region is not important for phylogenetic study. Therefore, we selected the D2-D3 region from the complete 28S rRNA sequences for phylogenetic analysis. Our analyses of 28S rRNA gene sequences showed that the 28S rRNA was useful as a phylogenetic marker for analyzing the relationships among Theileria spp. in ruminants. In addition, the D2-D3 region was a short segment that could be used instead of the whole 28S rRNA sequence during the phylogenetic analysis of Theileria, and it may be an ideal DNA barcode.

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Differentiation of two ovine Babesia based on the ribosomal DNA internal transcribed spacer (ITS) sequences

Cited by 45 publications

References 15 publications

Development of droplet digital PCR for the detection of Babesia microti and Babesia duncani

Development of droplet digital PCR for the detection of Babesia microti and Babesia duncani

Phylogeny of Theileria buffeli genotypes identified in the South African buffalo (Syncerus caffer) population

Phylogenetic Analysis of Ruminant Theileria spp. from China Based on 28S Ribosomal RNA Gene

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