Estabilshment Of Duplex PCR Assay For Detection MO And MA And An Indirect ELISA For Detection For MO

Mycoplasma ovipneumoniae (MO) is a kind of pathogenic bacteria which can cause the mycoplasmal pneumonia of sheep and goats (MPSG). The main clinical manifestations of infected sheep and goats are cough, gasp, gradual wasting and pathologic changes by hyperplastic inflammation of pulmonary interstitial tissue. The disease can bring huge economic losses to sheep and goats husbandry in China.In order to establish a duplex PCR method for simultaneous detection of MO and Mycoplasma arginini (MA), the research was based on designing the specific primers of MO and MA and optimizing the reaction conditions of PCR. Our laboratory have already been successfully expressed the hemolysin protein of MO. The recombinant protein was used as coating antigen to establish the indirect ELIS A to dectect the antibody of MO. The collected clinical samples were detected by the methods of bacteria cultivation, duplex PCR and indirect ELIS A, and the results were compared with each other.The bacteria cultivation results were concordant 81.37% of the duplex PCR. The duplex PCR was more sensitive than bacteria cultivation for the mycoplasma detection. The accordance ratio between the results of indirect ELISA and the bacteria cultivation was 94.85%. The detected results of cultivation and indirect ELISA were not significant (P>0.05).This study established fast, efficient and accurate detected pathogenic bacteria MO and MA methods which is containing the duplex PCR and indirect ELISA. And the study have practical significance to the rapid diagnosis of clinical pneumonia causing by MO.