细粒棘球绦虫半胱氨酸蛋白酶抑制剂基因的克隆、表达及诊断价值的初步评价

doi:10.11843/j.issn.0366-6964.2018.03.014

Abstract

Abstract:

This experiment was conducted to study the characterization and diagnostic value of cystatin of Echinococcus granulosus and provide basis for prevention and control of Echinococcus granulosus. We cloned and expressed Eg-cystatin, and presented a bioinformatic characterization, Western blotting. Immunofluorescence localization was performed to determine the distribution of cystatin from E. granulosus, and explore its potential for diagnosis of cystic echinococcosis (CE) in sheep based on indirect ELISA. The results showed that Eg-cystatin is belong to a typical type Ⅱ cystatin with an N-terminal signal peptide and the cystatin-like domain (G, Q-X-V-X-G, P-W). Phylogenetic analysis indicated that Eg-cystatin belongs to the cestode cystatin clade, and shares 72.20%-80.66% identity with cystatins from other cestodes. Native Eg-cystatin was located on the tegument and hooks of protoscoleces (PSCs), the whole germinal layer, and the inner body and eggs of adult worms. Indirect ELISA exhibited low specificity (79.1%) and sensitivity (83.3%), and the diagnostic accordance rate was 81.25% compared with the results of necropsy. Eg-cystatin was widely distributed in the larva and adult worm of E. granulosus, suggesting that it is closely related to the life activity of this parasite. However, rEg-cystatin is not a suitable antigen for the detection of sheep CE.

CLC Number:

S852.734

WU Mao-di, SONG Xing-ju, YAN Min, YANG Ai-guo, GUO Li, WANG Ning, XIE Yue, GU Xiao-bin, YANG Guang-you. Cloning, Expression and Preliminary Evaluation Diagnostic Value of Echinococcus granulosus Cystatin Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(3): 572-579.

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Cloning, Expression and Preliminary Evaluation Diagnostic Value of Echinococcus granulosus Cystatin Gene

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