Development of multiple-locus variable-number tandem-repeat analysis for rapid genotyping ofEhrlichia ruminantiumand its application to infectedAmblyomma variegatumcollected in heartwater endemic areas in Uganda

Abstract

SUMMARYThe rickettsial bacteriumEhrlichia ruminantiumis the causative agent of heartwater, a serious tick-borne disease in ruminants. The genetic diversity of organisms in the field will have implications for cross-protective capacities of any vaccine developed, and for an effective vaccine design strategy proper genotyping and understanding of existing genetic diversity in the field is necessary. We searched for variable-number tandem-repeat (VNTR) loci for use in a multi-locus VNTR analysis (MLVA). Sequencing analysis of 30 potential VNTRs using a panel of 17 reference strains from geographically diverse origins identified 12 VNTRs with allelic profiles differing between strains. Application of MLVA to 38E. ruminantium-infectedAmblyomma variegatumcollected from indigenous cattle in 6 different districts of Uganda identified 21 MLVA types. The discriminatory power of MLVA was greater than that ofmap1PCR-restriction fragment length polymorphism analysis, with which only 6 genotypes were obtained. The high discriminatory power as well as cost- effective performance of MLVA provide the potential for this technique to be applied in the future with respect to optimizing vaccine trials by identifying local strain diversity, and also raise the possibility of exploring the association betweenE. ruminantiumgenotypes and phenotypes such as pathological outcome in the ruminant host.