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Abstract: The diagnosis of piroplasms is achieved by staining methods such as Giemsa, feulgen or Methyl-green-puronin on blood smear or tick salivary gland. These methods are not specific and could be accompanied 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 as EDTA-blood, ethanol blood fixed, Giemsa
stained blood smear or from native were used for DNA extraction. Phenol/chloroform
extraction method, TriPure method and rapid DNA-Isolation method (Kit) were applied to compare DNA-isolation. Protozoan DNA could be amplified using extracted DNA from EDTA- blood with all three methods. DNA extracted from blood fixed in ethanol could be successfully 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 be demonstrated only when the DNA was extracted using rapid DNA-extraction method (kit). The results showed that the amount of blood for the PCR analysis revealed that less than 10 µl blood could be sufficient to detect protozoan parasites in the infected animals. These
results suggest that it is possible to 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 samples than the conventional methods. J.Vet.Res. 62,2:15-20,2007.