Recombinant multiepitope protein for early detection of dengue infections

被引:36
作者
AnandaRao, R
Swaminathan, S
Fernando, S
Jana, AM
Khanna, N
机构
[1] Int Ctr Genet Engn & Biotechnol, RGP Grp, New Delhi 110067, India
[2] Univ Sri Jayewardenepura, Fac Med Sci, Nugegoda, Sri Lanka
[3] Minist Def, Div Virol, Def Res & Dev Estab, Gwalior 474002, India
关键词
D O I
10.1128/CVI.13.1.59-67.2006
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
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 similar to 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.
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收藏
页码:59 / 67
页数:9
相关论文
共 26 条
[1]   A custom-designed recombinant multiepitope protein as a dengue diagnostic reagent [J].
AnandaRao, R ;
Swaminathan, S ;
Fernando, S ;
Jana, AM ;
Khanna, N .
PROTEIN EXPRESSION AND PURIFICATION, 2005, 41 (01) :136-147
[2]   Use of recombinant envelope proteins for serological diagnosis of dengue virus infection in an immunochromatographic assay [J].
Cuzzubbo, AJ ;
Endy, TP ;
Nisalak, A ;
Kalayanarooj, S ;
Vaughn, DW ;
Ogata, SA ;
Clements, DE ;
Devine, PL .
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, 2001, 8 (06) :1150-1155
[3]   PRECISE LOCATION OF SEQUENTIAL DENGUE VIRUS SUBCOMPLEX AND COMPLEX B-CELL EPITOPES ON THE NONSTRUCTURAL-1 GLYCOPROTEIN [J].
FALCONAR, AKI ;
YOUNG, PR ;
MILES, MA .
ARCHIVES OF VIROLOGY, 1994, 137 (3-4) :315-326
[4]   Recognition of synthetic oligopeptides from nonstructural proteins NS1 and NS3 of dengue-4 virus by sera from dengue virus-infected children [J].
Garcia, G ;
Vaughn, DW ;
DelAngel, RM .
AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 1997, 56 (04) :466-470
[5]  
GOEDDEL DV, 1990, METHOD ENZYMOL, V185, P3
[6]   Evaluation of six immunoassays for detection of dengue virus-specific immunoglobulin M and G antibodies [J].
Groen, J ;
Koraka, P ;
Velzing, J ;
Copra, C ;
Osterhaus, ADME .
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, 2000, 7 (06) :867-871
[7]   Cities spawn epidemic dengue viruses [J].
Gubler, DJ .
NATURE MEDICINE, 2004, 10 (02) :129-130
[8]   Dengue and dengue hemorrhagic fever [J].
Gubler, DJ .
CLINICAL MICROBIOLOGY REVIEWS, 1998, 11 (03) :480-+
[9]  
Gubler Duane J., 2002, Trends in Microbiology, V10, P100, DOI 10.1016/S0966-842X(01)02288-0
[10]   Potential effect of population and climate changes on global distribution of dengue fever: an empirical model [J].
Hales, S ;
de Wet, N ;
Maindonald, J ;
Woodward, A .
LANCET, 2002, 360 (9336) :830-834