Improved detection of Babesia bigemina from various geographical areas in Africa using quantitative PCR and reverse line blot hybridisation

dc.contributor.authorStoltsz, Wilhelm Heinrich
dc.contributor.authorByaruhanga, Charles
dc.contributor.authorTroskie, Milana
dc.contributor.authorMakgabo, Sekgota Marcus
dc.contributor.authorOosthuizen, Marinda C.
dc.contributor.authorCollins, Nicola E.
dc.contributor.authorDas Neves, Luis Carlos Bernardo G.
dc.date.accessioned2021-09-29T14:09:10Z
dc.date.available2021-09-29T14:09:10Z
dc.date.issued2020-07
dc.description.abstractBabesia bigemina is one of the aetiological agents of bovine babesiosis, which causes economic losses through mortality, loss of production and control costs. Effective means of detecting and quantifying B. bigemina in cattle populations is therefore important to inform control approaches. In order to examine the parasite genetic diversity in African countries, B. bigemina 18S rRNA genes from cattle from South Africa, Uganda and Angola were sequenced. The 25 distinct B. bigemina 18S rRNA gene sequences obtained in this study showed 99 to 100% identity with previously published sequences of strains from African and other continents. The sequences of the previously published B. bigemina 18S rRNA gene-specific quantitative PCR (qPCR) primers and probe, developed based on American and Asian strains, were conserved in the African B. bigemina sequences. The qPCR assay was evaluated using 10-fold and 2-fold serial dilutions of B. bigemina-infected erythrocytes to determine the efficiency and analytical sensitivity. The qPCR assay had an efficiency of 98.14 ± 1.71%, and the limit of detection was approximately 1.5 infected red blood cells (iRBCs) per microlitre (μl) of blood. The detection rate of B. bigemina from duplicates of field-collected blood samples from cattle from South Africa, Mozambique and Angola was 37% (30/81), 12% (6/49) and 50% (38/76), respectively. Reverse line blot hybridisation (RLB) results obtained from the same samples in previous studies, using a previously published B. bigemina-specific probe, detected the parasite DNA in only 1.5% (3/206) of the samples. A new B. bigemina-specific RLB oligonucleotide probe was designed in the hypervariable V4 region of the 18S rRNA gene. Screening of field blood samples from cattle showed that the new probe was specific, and its frequency of detection of B. bigemina was three times higher than the previously published probe. The qPCR assay and the newly developed B. bigemina-specific RLB probe provide good tools for epidemiological studies, which are essential in the control of bovine babesiosis.en_ZA
dc.description.departmentVeterinary Tropical Diseasesen_ZA
dc.description.librarianhj2021en_ZA
dc.description.sponsorshipThe Health and Welfare Sector Education and Training Authority (HWSETA), the Belgian Directorate General for Development Co-operation (DGCD) Framework agreement ITM/DGCD and the Claude Leon Foundation of South Africa.en_ZA
dc.description.urihttp://www.elsevier.com/locate/ttbdisen_ZA
dc.identifier.citationStoltsz, H., Byaruhanga, C., Troskie, M. et al. 2020, 'Improved detection of Babesia bigemina from various geographical areas in Africa using quantitative PCR and reverse line blot hybridisation', Ticks and Tick-borne Diseases, vol. 11, no. 4, art. 101415, pp. 1-9.en_ZA
dc.identifier.issn1877-959X (print)
dc.identifier.issn1877-9603 (online)
dc.identifier.other10.1016/j.ttbdis.2020.101415
dc.identifier.urihttp://hdl.handle.net/2263/82001
dc.language.isoenen_ZA
dc.publisherElsevieren_ZA
dc.rights© 2020 Elsevier GmbH. All rights reserved. Notice : this is the author’s version of a work that was accepted for publication in Ticks and Tick-borne Diseases. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting and other quality control mechanisms may not be reflected in this document. A definitive version was subsequently published in Ticks and Tick-borne Diseases, vol. 11, no. 4, art. 101415, pp. 1-9, 2020. doi : 10.1016/j.ttbdis.2020.101415.en_ZA
dc.subject18S rRNAen_ZA
dc.subjectPolymerase chain reaction (PCR)en_ZA
dc.subjectQuantitative PCR (qPCR)en_ZA
dc.subjectBabesia bigeminaen_ZA
dc.subjectDetectionen_ZA
dc.subjectDiagnosisen_ZA
dc.subjectReverse line blot (RLB)en_ZA
dc.titleImproved detection of Babesia bigemina from various geographical areas in Africa using quantitative PCR and reverse line blot hybridisationen_ZA
dc.typePostprint Articleen_ZA

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