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Recombinant multiepitope protein for early detection of dengue infections.
Clin Vaccine Immunol. 2006 Jan; 13(1):59-67.CV

Abstract

Dengue fever is a mosquito-borne viral disease prevalent mainly in tropical countries. As the clinical manifestations of dengue are not very unique, laboratory diagnosis is crucial in identifying cases of dengue infection. Detection of dengue infection based on the identification of antidengue antibodies has emerged as a practical and reliable means of diagnosing dengue fever. We recently developed a customized recombinant dengue multiepitope protein (r-DME-G) that can specifically detect the immunoglobulin G (IgG) class of antidengue antibodies in patient sera. Using this strategy, we have now created another dengue multiepitope protein, r-DME-M, with specificity for the IgM class of antidengue antibodies. A synthetic gene encoding the r-DME-M protein was expressed as a maltose-binding protein fusion in Escherichia coli. The recombinant protein was purified in a single affinity chromatographic step to obtain yields of approximately 15 mg purified protein/liter of culture. The purified protein was used to develop an in-house IgM enzyme-linked immunosorbent assay (ELISA) and tested using a panel of 172 patient sera characterized using the commercially available Dengue Duo rapid strip test from PanBio, Australia. The IgM ELISA results showed that the r-DME-M protein not only recognized all IgM(+) samples identified by the PanBio test but also identified samples missed by the latter test. We also successfully adapted the r-DME-M protein to a rapid strip test format. This approach of creating customized antigens coupled to overexpression in E. coli and simple purification offers a promising alternative option to dengue diagnosis with the potential to circumvent the drawbacks of the whole virus antigen-based commercial kits.

Authors+Show Affiliations

RGP Group, International Centre for Genetic Engineering and Biotechnology, P.O. Box 10504, Aruna Asaf Ali Marg, New Delhi-110067, India.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16426001

Citation

Anandarao, Ravulapalli, et al. "Recombinant Multiepitope Protein for Early Detection of Dengue Infections." Clinical and Vaccine Immunology : CVI, vol. 13, no. 1, 2006, pp. 59-67.
Anandarao R, Swaminathan S, Fernando S, et al. Recombinant multiepitope protein for early detection of dengue infections. Clin Vaccine Immunol. 2006;13(1):59-67.
Anandarao, R., Swaminathan, S., Fernando, S., Jana, A. M., & Khanna, N. (2006). Recombinant multiepitope protein for early detection of dengue infections. Clinical and Vaccine Immunology : CVI, 13(1), 59-67.
Anandarao R, et al. Recombinant Multiepitope Protein for Early Detection of Dengue Infections. Clin Vaccine Immunol. 2006;13(1):59-67. PubMed PMID: 16426001.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Recombinant multiepitope protein for early detection of dengue infections. AU - Anandarao,Ravulapalli, AU - Swaminathan,Sathyamangalam, AU - Fernando,Sirimali, AU - Jana,Asha M, AU - Khanna,Navin, PY - 2006/1/24/pubmed PY - 2006/4/14/medline PY - 2006/1/24/entrez SP - 59 EP - 67 JF - Clinical and vaccine immunology : CVI JO - Clin Vaccine Immunol VL - 13 IS - 1 N2 - Dengue fever is a mosquito-borne viral disease prevalent mainly in tropical countries. As the clinical manifestations of dengue are not very unique, laboratory diagnosis is crucial in identifying cases of dengue infection. Detection of dengue infection based on the identification of antidengue antibodies has emerged as a practical and reliable means of diagnosing dengue fever. We recently developed a customized recombinant dengue multiepitope protein (r-DME-G) that can specifically detect the immunoglobulin G (IgG) class of antidengue antibodies in patient sera. Using this strategy, we have now created another dengue multiepitope protein, r-DME-M, with specificity for the IgM class of antidengue antibodies. A synthetic gene encoding the r-DME-M protein was expressed as a maltose-binding protein fusion in Escherichia coli. The recombinant protein was purified in a single affinity chromatographic step to obtain yields of approximately 15 mg purified protein/liter of culture. The purified protein was used to develop an in-house IgM enzyme-linked immunosorbent assay (ELISA) and tested using a panel of 172 patient sera characterized using the commercially available Dengue Duo rapid strip test from PanBio, Australia. The IgM ELISA results showed that the r-DME-M protein not only recognized all IgM(+) samples identified by the PanBio test but also identified samples missed by the latter test. We also successfully adapted the r-DME-M protein to a rapid strip test format. This approach of creating customized antigens coupled to overexpression in E. coli and simple purification offers a promising alternative option to dengue diagnosis with the potential to circumvent the drawbacks of the whole virus antigen-based commercial kits. SN - 1556-6811 UR - https://www.unboundmedicine.com/medline/citation/16426001/Recombinant_multiepitope_protein_for_early_detection_of_dengue_infections_ L2 - http://cvi.asm.org/cgi/pmidlookup?view=long&pmid=16426001 DB - PRIME DP - Unbound Medicine ER -