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[Preparation of specific antibodies against recombinant Eml8 antigen and detection of the circulating antigen].
- Source :
-
Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases [Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi] 2006 Oct; Vol. 24 (5), pp. 329-32. - Publication Year :
- 2006
-
Abstract
- Objective: To prepare specific antibodies against the recombinant Eml8 antigen (ReEml8), establish a sandwich ELISA and detect the Eml8 circulating antigen in patients' sera.<br />Methods: Rabbits and BALB/c mice were immunized with the ReEml8 antigen, which was purified by affinity method for preparation of the specific poly-clonal and monoclonal antibodies. A sandwich ELISA was established by the specific antibodies.<br />Results: By immunizing with the ReEml8 antigen, high antibody level was reached with a serum dilution of 1: 204 800 and above in the immunized rabbits. After double selection by ELISA using the ReEml8 antigen and block ELISA using both AE-positive and negative control sera, 14 positive cell clones were obtained with an inhibition rate of more than 50%. Those mono-and poly-clonal antibodies were matched freely in sandwich ELISA tests for detecting the ReEml8 antigen. A combination of monoclonal antibody No.9 and polyclonal antibody showed the best result. The sensitivity to detect ReEml8 antigen was at 3 ng/ml. Six of 11 AE sera were positive when tested with the sandwich ELISA system.<br />Conclusion: Highly specific polyclonal and monoclonal antibodies have been prepared, and a sensitive sandwich ELISA established. Preliminary result is suggested that a detectable level of Eml8 circulating antigen is present in AE patients' sera.
- Subjects :
- Animals
Antigens, Helminth genetics
Antigens, Helminth immunology
Echinococcosis parasitology
Echinococcosis, Hepatic parasitology
Enzyme-Linked Immunosorbent Assay methods
Female
Humans
Immunization
Mice
Mice, Inbred BALB C
Rabbits
Recombinant Proteins immunology
Reproducibility of Results
Time Factors
Antibodies, Monoclonal blood
Antigens, Helminth blood
Echinococcosis blood
Echinococcosis, Hepatic blood
Echinococcus multilocularis immunology
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1000-7423
- Volume :
- 24
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases
- Publication Type :
- Academic Journal
- Accession number :
- 17361809