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Western Blot Detection of Human Anti-Chikungunya Virus Antibody with Recombinant Envelope 2 Protein
Authors:Zhaoshou Yang  Jihoo Lee  Hye-Jin Ahn  Chom-Kyu Chong  Ronaldo F Dias  Ho-Woo Nam
Institution:1Department of Parasitology, College of Medicine, Catholic University of Korea, Seoul 06591, Korea;2Department of Biochemistry, Chungbuk National University, Cheongju 28644, Korea;3Genbody Inc., Cheonan 31116, Korea;4Bahia Farma Diagnostics, Cep 43.700-000, Simoes, Bahia, Brazil
Abstract:Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection.
Keywords:Chikungunya virus (CHIKV)  envelope E2 protein  intrinsically unstructured domain (IUD)  GST-E2  MBP-E2  western blot  patient sera
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