Development Of A PCR Diagnostic Assay For E. Wenyoni Infection And Its Pathogeny Primary Biology Study

Eperythrozoonsis of cattle is a zoonosis caused by Eperythrozoon Wenyoni (E. Wenyoni). This disease hase been brought to the notice of people because of a large ecomonic losing in stockbreeding. In our country, E. Wenyoni has not been confirmed in molecule lever and its diagnostic assay has much shortcoming. This study has setted up the phylogenetic tree based on the 16SrRNA gene of E. wenyoni. In the same time, a polymerase chain reaction (PCR) diagnostic assay for E.wenyoni infection has been established.In order to confirm that our region exist E. wenyoni and research its classification place. Universal primers of 27F and 1492R were used to amplify the 16SrRNA gene of separated pathogeny that suspiciosed E. wenyoni by PCR. The results of cloning and sequencing showed that the segment comprises 1439bp, share 97.1% homology in bases with 16SrRNA gene of E. wenyoni that published in NCBI. The isolated E. wenyoni was named as E. wenyoni-CGX strain because of the sequence difference. The phylogenetic tree based on 16SrRNA gene indicates that the E. wenyoni and other Haemotrophic bacteria have formed a large phylogenenetic branch, which closes to the Mycoplasma genus of Mycoplasmatales (75%), while far from the pathogeny of Rickettsiales (55%). The results sustain the advice that the Eperythrozoon and haemobartonella genera should be reclassified in Mycoplasma genus of Mycoplasmatales.In order to establish a specificly, sensitively, fastly diagnostic assay for E.wenyoni infection, this study designed a pair genus specific primers based on the 16SrRNA gene of E.Wenyoni sequenced in our lab and a diagnostic PCR assay was established. The specific test and sensitive test showed that there was no cross reaction with Mycoplasma hyopneumoniae M. gallisepticum, E. coli, Salmonella enterica,...