Differentiation of sheep Theileria spp. and Babesia spp. By Polymerase Chain Reaction

The diagnosis of piroplasms is achieved by staining methods such as Giemsa, feulgen or Methylgreen-puronin on blood smear or tick salivary gland. These methods are not specific and could beaccompanied with some complications, especially in the carrier animals. In contrast to these methods,polymerase chain reaction is more sensitive and specific. Different sources of blood samples, such asEDTA-blood, ethanol blood fixed, Giemsa stained blood smear or from native were used for DNAextraction. Phenol/chloroform extraction method, TriPure method and rapid DNA-Isolation method(Kit) were applied to compare DNA-isolation. Protozoan DNA could be amplified using extractedDNA from EDTA- blood with all three methods. DNA extracted from blood fixed in ethanol could besuccessfully analyzed using Phenol/chloroform extraction method and rapid DNA extraction method(kit). But in the case of extracted DNA from native or stained blood smear, protozoan DNA could bedemonstrated only when the DNA was extracted using rapid DNA-extraction method (kit). The resultsshowed that the amount of blood for the PCR analysis revealed that less than 10 l blood could besufficient to detect protozoan parasites in the infected animals. These results suggest that it is possibleto analyze and control the already stained and registered blood smear from infected animals.Furthermore, the results facilitated possibility to develop simpler method for the collection of samplesthan the conventional methods. J.Vet.Res. 62,2:15-20,2007.

Differentiation of sheep Theileria spp. and Babesia spp. By Polymerase Chain Reaction Keywords:

Theileria , Babesia , diagnosis , DNA , PCR