Establishment Of PCR Diagnostic Method And Drug Screening In Vitro To Eperythrozoon Suis

Eperythrozoonosis, hematological disease infected by eperythrozoon parasitizing on the surface of RBC, in plasma and marrow causes decrease of hematokrit and concentration of hemoglobin and increase of leukocyte, anaemia, icterus and fever et cetera. Eperythrozoonosis is not only caused the enormous economic losses to animals,but also threaten the health of mankind seriously, and still there is not an effective method to diagnose, or effective drug to treat it. The purpose of this study is to offer theorial and technical supporting for diagnosis and treatment to eperythrozoonosis.In order to establish a special and sensitive method to diagnose eperythrozoonosis, the half-nest PCR primers were designed by Primer Premier5.0 software based on the special gene of eperythrozoon suis(AJ504999). According to analysis of the sequence in Gene Bank, we found that the homology between the fragment we cloned and the published sequence in Genebank is 100%, and there isn't homology between the fragment and other sequences published in Genebank. This DNA fragment hasn't been amplified from genome of streptococcus,anaplasma marginale, haemophilus parasuis, Chlamydia and theileria annulata.We identified 40 blood samples which collected from the hogpens around Lanzhou city, the positive rate of the special PCR diagnosis was 82.2%, which include portion of inapparent infection with no clinical symptoms.In order to screen effective medicines to treat eperythrozoonosis, some drugs which usually used for eperythrozoonosis in clinic and artesunate nano-emulsion which produced by our own lab were selected. Under the best culture condition in vitro, the effect of the drugs were judged by examination blood smears and calculation the infection rate of erythrocyte by using the microscopic. Furthermore, the concentration of some effective drugs were decided. It indicated that the effect of flavacridin hydrochloride was the best when the concentration was 500μg/mL, and it had less influence on erythrocytes. The effect of Artesunate Nano-emulsion is good, but the erythrocyte reputred in the process of culture. Berenil and doxycycline showed some certain role in eperythrozoon suis and other drugs were invalid to eperythrozoon suis.