Comparison of reverse line blot and -tubulin targeted nested polymerase chain reaction (PCR) techniques in the detection of Theileria and Babesia piroplasms in cattle in Uganda

Abstract

Three hundred (300) blood samples from apparently healthy cattle were collected in areas around Lake Mburo National Park in Western Uganda. These were used in the comparison of the sensitivity and specificity of reverse line blot (RLB) and β-tubulin targeted nested polymerase chain reaction (PCR) in the detection of tick-borne piroplasms in cattle. The sensitivity of RLB technique for the detection of Theileria andBabesia species was 71.25% (95% CI: 60.05 to 80.82%) and the specificity was 57.50% (95% CI: 40.89 to 72.960%) while the sensitivity of the beta-tubulin targeted nested PCR was 62.5% (95% CI: 50.96 to 73.08%) and the specificity was 72.5% (95% CI: 56.11 to 85.40%). The positive predictive value using RLB was 77.03% (95% CI: 65.79 to 86.01%) and the negative predictive value was 50% (95% CI: 34.90 to 65.10%) while the positive predictive value using β-tubulin targeted nested PCR was 81.97 % (95% CI: 70.02 to 90.64%) and the negative predictive value was 49.15% (95% CI: 35.89 to 62.50%). The Kappa statistic for level of agreement in detection of tick-borne piroplasms between RLB assay and β-tubulin targeted nested PCR was 0.7984 which indicated substantial agreement between the two tests. The RLB assay allowed the detection of individual species that simultaneously infected the cattle. However, it was not possible to identify the species with the β-tubulin targeted nested PCR. Key words: Babesia, Theileria, β-tubulin, reverse line blot, sensitivity, specificity.