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

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dc.contributor.author Stoltsz, Wilhelm Heinrich
dc.contributor.author Byaruhanga, Charles
dc.contributor.author Troskie, Milana
dc.contributor.author Makgabo, Marcus
dc.contributor.author Oosthuizen, Marinda C.
dc.contributor.author Collins, Nicola E.
dc.contributor.author Neves, L.C.B.G.D. (Luís)
dc.date.accessioned 2021-09-29T14:09:10Z
dc.date.available 2021-09-29T14:09:10Z
dc.date.issued 2020-07
dc.description.abstract Babesia 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.department Veterinary Tropical Diseases en_ZA
dc.description.librarian hj2021 en_ZA
dc.description.sponsorship The 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.uri http://www.elsevier.com/locate/ttbdis en_ZA
dc.identifier.citation Stoltsz, 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.issn 1877-959X (print)
dc.identifier.issn 1877-9603 (online)
dc.identifier.other 10.1016/j.ttbdis.2020.101415
dc.identifier.uri http://hdl.handle.net/2263/82001
dc.language.iso en en_ZA
dc.publisher Elsevier en_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.subject 18S rRNA en_ZA
dc.subject Polymerase chain reaction (PCR) en_ZA
dc.subject Quantitative PCR (qPCR) en_ZA
dc.subject Babesia bigemina en_ZA
dc.subject Detection en_ZA
dc.subject Diagnosis en_ZA
dc.subject Reverse line blot (RLB) en_ZA
dc.title Improved detection of Babesia bigemina from various geographical areas in Africa using quantitative PCR and reverse line blot hybridisation en_ZA
dc.type Postprint Article en_ZA


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