Preparation And Preliminary Application Of Monoclonal Antibodies Against Staphylococcus Aureus Enterotoxin B

Objective Monoclonal antibodies were prepared according to the protocol of hybridoma technique. It was aimed to investigate the prevalence of Staphylococcus aureus enterotoxins isolated from the hospital infections strains and SEB in Burn infections groups using ELISA which was newly developed by McAbs .Methods BALB/C Mice were immunized with purified Staphylococcus aureus enterotoxinB and McAbs against SEB were obtained according to hybridoma technique, Subsequently, A sandwich ELISA technique was developed and used to detect Staphylococcus aureus standard strain and hospital infections strains respectively, Furthermore to investigate the prevalence of Staphylococcus aureus enterotoxins and SEB in Burn infection groups.Results McAbs against SEB were obtained successfully,The titers of McAbs were between 1:12800 ~ 1:25600 in the pruified ascites ,The subclasses of these McAbs were IgG1 except IgG2(two strains)and the main immunological properties of these McAbs were Characterized, It showed that these McAbs had an identical epitopic specificity except two strains (3D6 and lDl).Cross-reactivity experiments showed that they had no reactivity with SEA, Escherichia coli Pseudomonas aeruginosa, Staphylococcus saprophticus andStreptococcus intestinalis. but had two strains cross-reactivity with SEC1 and one strain (polyclonal antibody).subsequenctly ,A11 McAbs recognized a 28.5KDa protein which was definded as SEB by Western-blot analysis. These data indicated that high titer of McAbs against SEB with high specificities and high stability were successfully established in this study.They also indicated these McAbs with high sensitivity (0.078 u g/L).The newly developed double sandwich ELISA could be used to detect SEB .These results showed the prevalence of SE in MRSA weresignificant(93.9%)compared with the MSSA group(28.6%).The prevalence of SE in SEB group(53.3%)were higher than SEC1 group(28.5%)and SEA(12.2%)group. Datas showed that SEB levels after burn followed by Staphylococcus aureus infection were significantly higher than those of normal controls and slight burn control group(p<0.01).Conclusions McAbs against SEB were successfully obtained. Subsequently ,A sandwich ELISA technique was developed and used to detect SEB .Datas showed this methods with high specificity n stability and sensitivity(0.078 U g/L) and could be used to detect SEB in hospital infection strains and in burn infection groups which might play an important role in the early diagnosis and cure of Staphylococcus aureus infection.