Design of enzyme-linked immunosorbent assay method for detection of anti-streptolysin-O antibodies on base of recombinant streptolysin-O

Ghasem Moosayebi; Hamid Abtahi; Ali Ghazavi; Nader zarinfar; Mohse Khaki; Mohammad Ali Payani

Koomesh

Journal of Semnan University of Medical Sciences

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Design of enzyme-linked immunosorbent assay method for detection of anti-streptolysin-O antibodies on base of recombinant streptolysin-O

Author(s):

Ghasem Moosayebi,

Hamid Abtahi

^,*,

Ali Ghazavi,

Nader zarinfar,

Mohse Khaki,

Mohammad Ali Payani

Koomesh:Vol. 13, issue 3; 362-368

Published online:May 25, 2012

Article type:Research Article

Received:Jan 02, 2011

Accepted:Sep 02, 2011

How to Cite:Ghasem MoosayebiHamid AbtahiAli GhazaviNader zarinfarMohse KhakiMohammad Ali Payaniet al.Design of enzyme-linked immunosorbent assay method for detection of anti-streptolysin-O antibodies on base of recombinant streptolysin-O.koomesh.13(3):e152522.

Abstract

Introduction: Group A, β hemolytic streptococci are among the major causative agents of otorhinolaryngology infections. Inadequate treatment of disease may lead to serious disorders such as acute rheumatic fever and glomerulonephritis. Anti-streptolysin-O (ASO) is commonly used as a marker in the diagnosis of infection. Purification of native streptolysin-O has several difficulties and its industrial production process is time consuming with very low yield and the risk of biological contamination. In this study, we used a recombinant streptolysin-O protein as an antigen to detect ASO antibodies in enzymes-linked immunosorbent assay (ELISA). Materials and Methods: We amplified streptolysin-O gene by polymerase chain reaction (PCR) method and subcloned in prokaryotic expression vector PET28a. E.coli. BL21-DE3-plySs strain was transformed with PET28a-streptolysin-O and gene expression was induced by IPTG. ELISA microplates were coated with different concentration of streptolysin-O protein. Level of ASO antibodies were detected by ELISA method. The results obtained from ELISA method were compared with inhibition of hemolysis assay as a standard method. Results: The results showed that there is a positive significant correlation between the ELISA and inhibition of hemolysis method(r=0.97, p=0.0001). The sensitivity and specificity of ELISA for detection of ASO anti bodies were 100% and 83%, respectively. Conclusion: ELISA developed with recombinant streptolysine O showed a good sensitivity for detection of ASO antibodies. It is suggested that this method could be suitable for immunoassays.

Keywords

Streptococci pyogene, Recombinant streptolysin-O, Anti-streptolysin-O antibodies, Enzyme-linked immunosorbent assay

استرپتوکک پیوژن، استرپتولیزین-O نوترکیب، الایزا، آنتی‌بادی ضد استرپتولیزین-O

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