Prokaryotic expression and immunofluorescence localization of nucleoside diphosphate kinase gene from Dirofilaria immitis

doi:10.3969/j.issn.1004-1524.2019.09.08

›› 2019, Vol. 31 ›› Issue (9): 1453-1460.DOI: 10.3969/j.issn.1004-1524.2019.09.08

• Animal Science • Previous Articles Next Articles

Prokaryotic expression and immunofluorescence localization of nucleoside diphosphate kinase gene from Dirofilaria immitis

ZHANG Haojie¹, LIU Mei¹, LI Chunyan¹, HE Ran¹, LAN Jingchao², LUO Li², GU Xiaobin¹, XIE Yue¹, YANG Guangyou^1,*

1. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;
2. Chengdu Research Base of Giant Panda Breeding, Chengdu 610081, China

Received:2019-03-14 Online:2019-09-25 Published:2019-10-11

Abstract

Abstract: In order to provide base of heartworm diagnose, this study analyzed the molecular characteristics and identified the diagnostic potential of Dirofilaria immitis nucleoside diphosphate kinase (NDPK) gene. Di-NDPK was cloned and expressed, and the protein was analyzed by bioinformatic characterization, Western blotting and immunohistochemical analysis. Also the diagnostic potential of rDi-NDPK was evaluated by indirect ELISA. The results of Western blotting indicated that Di-NDPK had good reactivity. Immunohistochemical analysis results showed that Di-NDPK was distributed in hypodermis and intestinal epithelial cells of D.immitis. The indirect ELISA results showed that the sensitivity and specificity were 66.7% (16/24) and 38.9% (14/36) respectively. The cross reactions were observed with serum samples positive for Echinococcus granulosus, Ancylostoma caninum and Ascaris alata. In conclusion, Di-NDPK might be a candidate vaccine antigen, but not a suitable antigen for the detection of D. immitis.

Key words: Dirofilaria immitis, nucleoside diphosphate kinase gene, clone, prokaryotic expression, immunohistochemical analysis

CLC Number:

S855.99

ZHANG Haojie, LIU Mei, LI Chunyan, HE Ran, LAN Jingchao, LUO Li, GU Xiaobin, XIE Yue, YANG Guangyou. Prokaryotic expression and immunofluorescence localization of nucleoside diphosphate kinase gene from Dirofilaria immitis[J]. , 2019, 31(9): 1453-1460.

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Prokaryotic expression and immunofluorescence localization of nucleoside diphosphate kinase gene from Dirofilaria immitis

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