Reverse Line Blot Hybridization with Species-Specific Oligonucleotide Probes: Application to Piroplasm Detection

Hurtado, Ana

doi:10.1007/978-1-4939-2004-4_14

Cited by 6 publications

(3 citation statements)

References 16 publications

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“…Alternative molecular methods include reverse line blot, which allows simultaneous screening of large numbers of pathogens using probe hybridization (Gubbels et al 1999, Hurtado 2015, and modern high-throughput screening methods such as the microfluidic real-time PCR system (Fluidigm) (Michelet et al 2014) and Next Generation sequencing (Bonnet et al 2014).…”

Section: Pcr Analysismentioning

confidence: 99%

Guidelines for the Detection ofBabesiaandTheileriaParasites

Lempereur

Beck

Fonseca

et al. 2017

Vector-Borne and Zoonotic Diseases

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The genera Babesia and Theileria (phylum Apicomplexa, order Piroplasmida) are mainly transmitted by Ixodid ticks in which the sexual part of their life cycle followed by sporogony takes place. They include protozoan parasites that infect erythrocytes of a variety of vertebrate hosts, including domestic and wild animals, with some Babesia spp. also infecting humans. Babesia sporozoites transmitted in the tick's saliva during the bloodmeal directly infect erythrocytes, where they asexually multiply to produce pear-shaped merozoites in the process of merogony; whereas a pre-erythrocytic schizogonic life stage in leukocytes is found in Theileria and precedes merogony in the erythrocytes. The wide spectrum of Babesia and Theileria species and their dissimilar characteristics with relation to disease severity, transmission, epidemiology, and drug susceptibility stress the importance of accurate detection of babesiosis and theileriosis and their causative agents. These guidelines review the main methods currently used for the detection of Babesia and Theileria spp. for diagnostic purposes as well as epidemiological studies involving their vertebrate hosts and arthropod vectors. Serological methods were not included once they did not indicate current infection but rather exposure.

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Section: Pcr Analysismentioning

confidence: 99%

Guidelines for the Detection ofBabesiaandTheileriaParasites

Lempereur

Beck

Fonseca

et al. 2017

Vector-Borne and Zoonotic Diseases

View full text Add to dashboard Cite

show abstract

“…Alternative molecular methods include reverse line blot, which allows simultaneous screening of large numbers of pathogens using probe hybridization , Hurtado 2015, and modern high-throughput screening methods such as the microfluidic real-time PCR system (Fluidigm) and Next Generation sequencing (Bonnet et al 2014).…”

Section: Pcr Analysismentioning

confidence: 99%

European Network for Neglected Vectors and Vector-Borne Infections COST Action Guidelines: What Is This About and What Is This For?

Charrel

Lempereur

Mihalca

et al. 2017

Vector-Borne and Zoonotic Diseases

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“…Reverse line blot hybridization (RLB) test is also used to simultaneously detect DNA from different piroplasm in blood or tick samples. Although this technique is technically elaborate, it has been applied to a high number of epidemiological studies in cattle, small ruminants, and ticks around the world (Hurtado 2015;Schötta et al, 2017).…”

Section: Diagnosismentioning

confidence: 99%

PCR-based detection and differentiation of piroplasms and malaria parasites from goats in Thailand

Huynh

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A novel multiplex PCR (mPCR) was developed to identify and differentiate malaria parasite (Plasmodium caprae),�and two piroplasms (Babesia bigemina�and�Theileria luwenshuni)�in Thai goats. Three pairs of primer were designed to be species-specific,�targeting cytochrome c oxidase subunit I (COI) gene in mitochondria.�Plasmids containing each parasite's DNA was serially diluted to evaluate the sensitivity of singleplex and mPCR assays. The specificity of the primer sets was confirmed by testing for amplification from DNA of each pathogen and other related hemoparasites. A total of 100 goat blood samples from five provinces of Thailand was used to validate the reliability and application of the assay. The detection limit of the mPCR in this study was 108 parasite copies and the diagnostic specificity in detection of P. caprae and T. luwenshuni was found to be high (94.9%-100%). The high level of diagnostic sensitivity (100%) in the detection of T. luwenshuni was also recorded. However, the diagnostic sensitivity of mPCR was found to be low (50%) in the detection of P. caprae from field-collected blood samples. These results demonstrated the successful establishment of the mPCR presented by�its same level of sensitivity to each single PCR counterparts and its ability to simultaneous detection all three parasites�in one reaction. This study also describes the first report of�Theileria luwenshuni�and�Babesia bigemina�in goats of Thailand.

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Reverse Line Blot Hybridization with Species-Specific Oligonucleotide Probes: Application to Piroplasm Detection

Cited by 6 publications

References 16 publications

Guidelines for the Detection ofBabesiaandTheileriaParasites

Guidelines for the Detection ofBabesiaandTheileriaParasites

European Network for Neglected Vectors and Vector-Borne Infections COST Action Guidelines: What Is This About and What Is This For?

PCR-based detection and differentiation of piroplasms and malaria parasites from goats in Thailand

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