Multiplex hydrolysis-probe assay for the simultaneous detection of Theileria equi and Babesia caballi infections in equids

Bhoora, Raksha; Pienaar, Ronel; Cornelius, Fran; Josemans, Antoinette I.; Matthee, Olivier; Marumo, Ratselane; Troskie, Christo; Mans, Ben J.

doi:10.1016/j.vetpar.2018.03.022

Cited by 13 publications

(15 citation statements)

References 25 publications

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“…These methods can also be designed to distinguish between parasite species or genotypes. Currently, these methods are more often used for research than in clinical practice [2,3,5,20,68,[85][86][87][88][89][90][91][92]. Numerous assays targeting one or multiple EP parasites, including conventional PCR [86,87], nested PCR (nPCR) [90,93], real-time PCR (rtPCR) [82,[94][95][96][97][98][99], multiplex PCR (mPCR) [87,89], reverse line blot (RLB) [100,101], and loop mediated isothermal amplification (LAMP) [85,88,91], have been developed.…”

Section: Diagnosismentioning

confidence: 99%

“…Currently, these methods are more often used for research than in clinical practice [2,3,5,20,68,[85][86][87][88][89][90][91][92]. Numerous assays targeting one or multiple EP parasites, including conventional PCR [86,87], nested PCR (nPCR) [90,93], real-time PCR (rtPCR) [82,[94][95][96][97][98][99], multiplex PCR (mPCR) [87,89], reverse line blot (RLB) [100,101], and loop mediated isothermal amplification (LAMP) [85,88,91], have been developed. Several of these assays were determined to have high sensitivity, with a detection limit of 10 −7 % parasitized erythrocytes (PE) [2,3,5,20,68,[85][86][87][88][89][90][91][92].…”

Section: Diagnosismentioning

confidence: 99%

“…Numerous assays targeting one or multiple EP parasites, including conventional PCR [86,87], nested PCR (nPCR) [90,93], real-time PCR (rtPCR) [82,[94][95][96][97][98][99], multiplex PCR (mPCR) [87,89], reverse line blot (RLB) [100,101], and loop mediated isothermal amplification (LAMP) [85,88,91], have been developed. Several of these assays were determined to have high sensitivity, with a detection limit of 10 −7 % parasitized erythrocytes (PE) [2,3,5,20,68,[85][86][87][88][89][90][91][92]. Quantitative methods, such as rtPCR (qPCR), have also been developed, but are mostly applied to increase the sensitivity of parasite detection, and are rarely used to evaluate parasite loads [94][95][96][97][98][99]102].…”

Section: Diagnosismentioning

confidence: 99%

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Twenty Years of Equine Piroplasmosis Research: Global Distribution, Molecular Diagnosis, and Phylogeny

et al. 2020

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Equine piroplasmosis (EP), caused by the hemoparasites Theileria equi, Theileria haneyi, and Babesia caballi, is an important tick-borne disease of equines that is prevalent in most parts of the world. Infection may affect animal welfare and has economic impacts related to limitations in horse transport between endemic and non-endemic regions, reduced performance of sport horses and treatment costs. Here, we analyzed the epidemiological, serological, and molecular diagnostic data published in the last 20 years, and all DNA sequences submitted to GenBank database, to describe the current global prevalence of these parasites. We demonstrate that EP is endemic in most parts of the world, and that it is spreading into more temperate climates. We emphasize the importance of using DNA sequencing and genotyping to monitor the spread of parasites, and point to the necessity of further studies to improve genotypic characterization of newly recognized parasite species and strains, and their linkage to virulence.

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Section: Diagnosismentioning

confidence: 99%

Section: Diagnosismentioning

confidence: 99%

Section: Diagnosismentioning

confidence: 99%

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Twenty Years of Equine Piroplasmosis Research: Global Distribution, Molecular Diagnosis, and Phylogeny

et al. 2020

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“…Determining the minimum parasite dose required by the vectors in South Africa to transmit these parasites does not fall within the scope of this study and therefore a tick transmission study was not conducted. However, we could infer that based on the 95% sensitivity of the T. equi qPCR assay (Bhoora et al, 2018), at a cut-off C q value of 37, the multiplex EP assay could detect as little at 10 5.1 parasites/ml. Given that the observed C q values of T. equi detected in zebra, fall below the reported cut-off C q of 37, we can conclude that the observed parasitaemia is zebra could allow for successful transmission by the tick vectors.…”

Section: Discussionmentioning

confidence: 99%

“…The presence of T. equi and B. caballi was detected using the multiplex EP real-time PCR (qPCR) assay as described by Bhoora et al (2018). The samples were run using Luna ® Universal qPCR Master Mix (New England Biolabs) as per manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Translocation a potential corridor for equine piroplasms in Cape mountain zebra (Equus zebra zebra)

Smith

Bhoora

Kotzé

et al. 2019

International Journal for Parasitology: Parasites and Wildlife

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Translocation of animals in fragmented habitats is an important means of dispersal and gene flow, however, the movement of animals has led to the spread of various diseases globally and wildlife are often the reservoirs of these diseases. Currently, Cape mountain zebra are translocated within South Africa as a management method for augmentation of isolated and fragmented populations. The movement of pathogens due to translocations in local regions have gone largely unchecked, particularly where there may still be isolated regions that can be negatively affected. Equine piroplasmosis is a tick-borne disease caused by Theilaria equi and/or Babesia caballi reported to occur in equids (Bhoora et al., 2010; Zweygarth et al., 2002). Here, the presence of T. equi and B. caballi was detected in 137 clinically healthy Cape mountain zebra from three South African reserves, Mountain Zebra National Park (MZNP), De Hoop Nature Reserve (DHNR) and Karoo National Park (KNP) using the multiplex EP real-time PCR (qPCR) assay. We observed 100% prevalence for T. equi and identified only one animal from MZNP with B. caballi. These results affirm that precautions should be taken prior to founding new populations of Cape mountain zebra and that potential farms and properties adjacent to prospective reserves should be screened for the presence of the organisms in order to mitigate risks of infection to domestic animals.

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Comparison of PCR-based methods for the detection of Babesia caballi and Theileria equi in field samples collected in Central Italy

et al. 2021

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Multiplex hydrolysis-probe assay for the simultaneous detection of Theileria equi and Babesia caballi infections in equids

Cited by 13 publications

References 25 publications

Twenty Years of Equine Piroplasmosis Research: Global Distribution, Molecular Diagnosis, and Phylogeny

Twenty Years of Equine Piroplasmosis Research: Global Distribution, Molecular Diagnosis, and Phylogeny

Translocation a potential corridor for equine piroplasms in Cape mountain zebra (Equus zebra zebra)

Comparison of PCR-based methods for the detection of Babesia caballi and Theileria equi in field samples collected in Central Italy

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