Observation Of Eperythrozoon Canis Morphology And Establishment Of PCR For Eperythrozoon Canis

Eperythrozoon Canis disease is Eperythrozoon attached the red cell membranes,bonemarrow and blood plasma of canine and causing canine anemia, fever, jaundice, and progressiveweight loss and other symptoms. Eperythrozoon Canis disease is a zoonotic ill, seriouslyhampered the development of canine breeding in China. Currently the examination of this pathgenin clinical veterinary mainly through microscopic, including the method of blood pressure andblood smear piece, but with more misdiagnosis of Eperythrozoon Canis. So,it is very necessary toestablish an efficient and specific detection methods for clinical diagnosis. This research includestwo aspects: The observation of Eperythrozoon canis morphology and the establishment of PCRfor Eperythrozoon canis. Morphological research was mainly carried through the collection andpreparation of fresh blood from the infected dogs,ultrathin sections were observed used the TEM.After the research of morphology in Eperythrozoon canine,the results suggested that theEperythrozoon canine could attache to the surface of red blood cells,or available free in theplasma, with pleomorphic, rod-shaped, comma-shaped ring, both contained RNA and DNAstructure; with the diameter of03to20μm, with a single limiting membrane circular structurewrapped parcel ribosome, no obvious organelles and nuclear-like structure, typical of prokaryotes.After the treatment used intramuscular injections of oxytetracycline and gave supportive therapyof clinical symptomatic treatment according to individual circumstances and different infections,we achieved good therapeutic effect, the cure rate is about80%, using this method can effectivelyreduce the mortality rate of infection in dogs. In other hand, according to16S rRNA genesequences of Eperythrozoon different species published in GenBank,the specific primers weredesigned from conserved regions and established the PCR diagnostic methods of Eperythrozooncanis, the results show that this method can be amplified530bp fragment from positive bloodsample, consistent with the expected results,and after the specific experimental validation of thismethod could only amplified fragment from dogs Eperythrozoon consistent with the purpose ofthe strip, the blood of healthy dogs, canine Toxoplasma gondii, canine parvovirus virus all withno fragment.The sensitivity experiments of this method suggested that this method is only8pg.With this method we deteced the blood samples from clinical cases,the results we got11positive samples from15suspected positive blood samples with the rate of about73%and got5positivesamples from26Wright’s staining were negative with the rate of20%, were finally diagnosed asinfected Eperythrozoon.So,this study confirmed that the PCR method was accurated and better fordetecting the Eperythrozoon canis compared with Wright staining. The PCR method withadvantages of rapid, specificity, sensitivity and so on,could be used for diagnosis andepidemiological investigation of Eperythrozoon canis, and for understanding the etiology andepidemiology, clinical treatment and prevention.