| Eperythrozoonosis is one of zoonosis which is caused by attaching to the red blood cells of multiple warm-blooded animals with Eperythrozoon.It still has great controversy on its biological classification,in traditionally biological classification it was sorted as rickettsia,eperythrozoon.In recent years,molecular classification results show that it is closer to mycoplasma,and named it as hemotropic mycoplasma(Hemoplasma).The main clinical signs involve in high fever,anemia,jaundice,and decreasing of reproductive performance.To Eperythrozoon.suis can cause the declining of the sow's reproductive ability and decreasing of the immunity of the piglet.Eperythrozoon is a kind of prokaryote,usually its shape is spherical,but no cell wall,only has single cell membrane,generally there is 1-2 on surface of red blood cells,more up to 6-7,which can lead to deformation of the parasited red blood cells with star or irregular shape.Among these diagnose measures,the microscope examination is the most commonly used including fresh blood pressure and smear staining method,but it is taken effect by the operator ability and blood osmotic pressure inside and outside,so it often easily to be caused false positive.The methods of serological diagnosis involve in Indirect hemagglutination assay(IHA),Ensyme-linked immunosorbent assay(ELISA)and complement fixation test(CFT),because the change of antibody levels is no regularity or no steadiness during the height of bacillemia,it easily cause false negative by serological examination.Given these circumstances,it is necessary to establish a rapid,specific,sensitive detection method to diagnose and control the disease.This study first conducted the establishment of moleculardetection for E.suis According to the eported 16 S rRNA gene sequence from Gen bank(U88565),the specific primers was designed,then the blood genomic DNA was extracted,the conventional PCR amplification was applied to determined it,meanwhile its specificity tests and sensitivity test also was made.During from January 2016 to December 2016,all 155 blood samples were collected from Hefei city,Wuhu city,Fuyang city,Huangshan city,then they were detected by the established PCR method to take epidemiology investigation,and the blood smear stain also was taken to confirm coincidence rate.The obtained data was analyzed including season,feeding,etc.The results showed that:(1)the established PCR specifically could amplify the aim gene,its size was 703 bp.The sequence homology displayed that it highly was simarity to the reported mycoplasma(FN436012.1),the homology was 94.9%.Its specificity was ideal,it can't amplify Babesia bovis,Babesia bigemina,Babesia canis,Streptococcus,Pasteurella,Toxoplasma gondii and Theileria ovis.Additionally,its sensitivity also was better,the minimum amount of DNA for detection was 0.25 ?g.(2)Epidemiological investigation showed that the detection rate of 155 samples from Anhui province was 69.68%,but the detection rate was only 40.64% by Giemsa Staining,its detection rate was higher than that of Giemsa Staining.Statistics analysis showed that the positive rate was 73.91% in spring and summer season,but it was lower in the autumn and winter,the positive rate only is 66.28%;In month age,the positive detection rate of the piglets was the highest in 1-3 months,it indicated that the piglet was more susceptible to the disease.In a conclusion,it still was more serious at some pig farmers in Anhui,especially in the spring and the summer,it maybe could be transformed by insect.The suggestion was given that it should arise to the awareness of the disease and improve the administration measures to prevent and control it. |
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