鸭坦布苏病毒E蛋白特异性抗原表位鉴定及Epitope-ELISA血清学诊断方法建立

doi:10.11843/j.issn.0366-6964.2017.10.019

Abstract

Abstract:

In this study, a unique antigenic epitope ₈₇YAEYI₉₁on E protein of duck Tembusu virus (DTMUV) was identified by phage display technology with purified monoclonal antibodies (MAb) 1A5 against DTMUV E protein. To determine whether the ₈₇YAEYI₉₁sequence is conserved among the E proteins of flaviviruses, we aligned the E protein amino acid sequences of flaviviruses. The results revealed that YAEYI was highly conservative in DTMUV strain, but the epitope sequence wasn't conserved among the corresponding amino acid sequence of the E protein in other strains of flavivirus. Dot blotting assay showed that epitope ₈₇YAEYI₉₁ had a good affinity with positive sera to DTMUV, and there was not cross-reacting with JEV-, DENV-, WNV-positive sera, suggesting that ₈₇YAEYI₉₁ was DTMUV unique epitope. An Epitope-ELISA assay was established by using unique epitope (₈₇YAEYI₉₁) fused protein and used to test duck serum samples which were also detected by the Neutralization Test as the standard method. According to results, the specificity of this test was 100%. The sensitivity of this epitope-based peptide serologic test for DTMUV infection was 96%. So, Epitope-ELISA was a specific method for detecting antibodies against DTMUV.

CLC Number:

S852.659.6

LI Chen-xi, BAI Xiao-fei, SHAOZHOU Wu-lin, ZHANG Qing-shan, LIU Ming, ZHANG Yun. Unique Antigenic Epitope Identification of Duck Tembusu Virus Envelope Protein and Development of Epitope-ELISA[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(10): 1958-1968.

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Unique Antigenic Epitope Identification of Duck Tembusu Virus Envelope Protein and Development of Epitope-ELISA

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