Volume 46, Issue 2 (6-2022)
Research in Medicine 2022, 46(2): 30-39 |
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Ethics code: IR.PII.REC.1399.006
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Hejazi M, Paryan M, Shokri R. Design and development of ELISA technique for detection of IgG against Neisseria meningitidis using purification of antisera. Research in Medicine 2022; 46 (2) :30-39
URL: http://pejouhesh.sbmu.ac.ir/article-1-3026-en.html
URL: http://pejouhesh.sbmu.ac.ir/article-1-3026-en.html
Department of Antigen and Antisera production, Production and Research Complex, Pasteur Institute of Iran, Karaj, Iran. , mparyan@gmail.com
Abstract: (1826 Views)
Background and Aim: Neisseria meningitidis is a leading cause of death in children in developed countries. Meningococcus has 13 sero-groups based on the capsule antigens. Rapid diagnosis of the bacteria results in timely selection of the right anti-microbial treatment. The present study, conducted in 2019, aimed to produce a highly pure antiserum for diagnostic assays for the detection of meningococcus sero-groups.
Methods: The type of research study was research and development. Inactivated bacterial suspension was intravenously injected into white New Zealand Rabbits. Serum was collected after the last injection. Purification process was performed using precipitation, chromatography, and tangential flow filtration. The produced antiserum was used in Agglutination and ELISA assays. The purified antiserum had a purity of more than 98% at a suitable quantity. Agglutination test with the primary antigen showed 4+ result. To develop the ELISA assay, the optimized concentration of primary and biotinylated antibodies was 1 and 80 micrograms, respectively. The results of positive predictive value were 100% and negative predictive value was 96%, for the designed assay.
Results: The purified antiserum had a purity of more than 98% at a suitable quantity. Agglutination test with the primary antigen showed 4+ result. To developed the ELISA assay, the optimized concentration of primary and biotinylated antibodies was 1 and 80 micrograms, respectively. The results of positive predictive value were 100% and Negative predictive value was 96%, for the designed assay.
Conclusion: Serum-based assays are recommended for the timely diagnosis of the disease since these assays are specific, sensitive, inexpensive, and rapid. The antiserum produced in the present research is highly pure. Therefore, we recommend the antiserum for the development of agglutination and ELISA assays for the detection of N. meningitides.
Methods: The type of research study was research and development. Inactivated bacterial suspension was intravenously injected into white New Zealand Rabbits. Serum was collected after the last injection. Purification process was performed using precipitation, chromatography, and tangential flow filtration. The produced antiserum was used in Agglutination and ELISA assays. The purified antiserum had a purity of more than 98% at a suitable quantity. Agglutination test with the primary antigen showed 4+ result. To develop the ELISA assay, the optimized concentration of primary and biotinylated antibodies was 1 and 80 micrograms, respectively. The results of positive predictive value were 100% and negative predictive value was 96%, for the designed assay.
Results: The purified antiserum had a purity of more than 98% at a suitable quantity. Agglutination test with the primary antigen showed 4+ result. To developed the ELISA assay, the optimized concentration of primary and biotinylated antibodies was 1 and 80 micrograms, respectively. The results of positive predictive value were 100% and Negative predictive value was 96%, for the designed assay.
Conclusion: Serum-based assays are recommended for the timely diagnosis of the disease since these assays are specific, sensitive, inexpensive, and rapid. The antiserum produced in the present research is highly pure. Therefore, we recommend the antiserum for the development of agglutination and ELISA assays for the detection of N. meningitides.
Type of Study: Applicable |
Subject:
Immunology
Received: 2021/03/16 | Accepted: 2021/12/1 | Published: 2022/08/18
Received: 2021/03/16 | Accepted: 2021/12/1 | Published: 2022/08/18
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