A review of nucleic-acid-based diagnostic tests for Babesia and Theileria, with emphasis on bovine piroplasms.

Nucleic acid-based methods offer a variety of tools for the detection of parasites. This field of veterinary and medical sciences is rapidly evolving, leading to greater sensitivity and higher throughput. One of the reasons justifying such a fast development is the fact that tests targeting several taxa can be created. The present article deals with the applications of molecular diagnostics of tick-borne diseases in Parasitology. Special attention is given to Babesia and Theileria species infecting livestock. The commonly used molecular methods in diagnostic of tick-transmitted hematic protozoa are the following: (i) final time polymerase chain reaction; (ii) reverse line blotting (RLB); (iii) real time PCR, based on SYBR Green or probe fluorescence; (iv) isothermal amplification methods: loop-mediated amplification (LAMP) and self sustaining sequence replication (3SR, also named as "Nucleic Acid Sequence Based Amplification", NASBA, or Transcription Mediated Amplification, TMA). In general, none of these methods could be considered better than another. Their score in diagnostic applications greatly depends on the laboratory size. Small-scale laboratories handling few samples may find final time PCR a cheap alternative. On the contrary, large-scale laboratories prefer methods amenable to automation, like RLB, PCR-ELISA or qPCR.