Production of IgM Specific Recombinant Dengue Multiepitope Protein for Early Diagnosis of Dengue Infection

详细信息    查看全文

• 作者：Nagesh K. Tripathi ; Ambuj Shrivastva ; Priyabrata Pattnaik ; Manmohan Parida ; Paban K. Dash ; Nimesh Gupta ; Asha M. Jana ; P. V. Lakshmana Rao
• 刊名：Biotechnology Progress
• 出版年：2007
• 出版时间：April 2007
• 年：2007
• 卷：23
• 期：2
• 页码：488 - 493
• 全文大小：66K
• 年卷期：v.23,no.2(April 2007)
• ISSN：1520-6033

文摘

Dengue virus infections have recently undergone dramatic expansion in range, affecting severaltropical and subtropical regions of the world. Early detection of dengue infection based on theidentification of antibodies has emerged as a practical and reliable means of diagnosis of denguefever. The recombinant dengue multiepitope (rDME-M) protein specific to IgM in E. coli wasproduced in a 5-L fermentor for use in diagnostic purpose. After fermentation, dry cell weightwas approximately 11.8 g/L of the culture. The rDME-M protein was purified under denaturingconditions using single-step nickel nitrilotriacetate (Ni-NTA) affinity chromatography. The finalyield of purified rDME-M protein from this method was approximately 68.5 mg/L of the culture.The purity of rDME-M protein was checked by SDS-PAGE analysis, and the reactivity of thisprotein was further checked by Western blotting and enzyme-linked immunosorbent assay(ELISA). The purified protein was used as an antigen in the development of an in-house dipstickELISA and evaluated with a panel of 80 patient sera, characterized using commercially availabletests for detection of dengue antibody. The results were in excellent agreement with those ofIgM capture ELISA (Pan-Bio) and rapid immunochromatography (IC) test (Pan-Bio). Theseresults show that the in-house dipstick ELISA using rDME-M protein can be used as a promisingkit because of its comparable sensitivity, specificity, field applicability, and low cost.