| Brucella is an intracellular,gram-negative coccobacillus,and the zoonotic disease caused by Brucella is known as brucellosis.Both Omp25 and Omp31 are the group 3 of outer membrane proteins(OMPs)of Brucella.Omp25 can bind to special epitopes that other outer membrane proteins cannot recognize,and Omp31 is an effective protective antigen.Both proteins have the functions of enhancing humoral and cellular immunity.Therefore,it is of great practical significance to establish a serological detection and diagnosis method with Omp25 and Omp31 as coating antigens.Among the various methods for diagnosing brucellosis,the experimental equipment and environmental requirements for serological testing are easier to achieve,which makes serological testing have advantages over bacteriological testing and molecular biological testing.In this study,to establish a serological method for the early and rapid diagnosis of brucellosis,and further strengthen the prevention and control of brucellosis,we first obtained recombinant purified Omp31(B.melitensis)and Omp25(B.ovis).Secondly,an indirect ELISA method using Omp25 and Omp31 proteins as coating antigens was established.It has been verified that the detection results of this method are in 100% agreement with the results of the the Rose Bengal test.Then,100 unknown sheep sera from Shu'lan,Nong'an,Da'an,Tong'yu and De'hui were detected by this method.The results suggest that the positive detection rate was(82/100)when the coating antigen was Omp25,and the positive detection rate was(76/100)when the coating antigen was Omp31.The results indicate that the recombinant expressed Omp25 and Omp31 have good antigenicity,and the established indirect ELISA method has good stability and sensitivity.Besides,BLI technology was used to screen the nucleic acid aptamer of the Omp31 protein of the outer membrane protein of Brucella sheep,which would provide a basis and a new strategy for aptamers detection of brucellosis in the future. |
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