基于狂犬病病毒重组蛋白的ELISA检测方法比较研究

Abstract

Abstract:

This study was designed to compare the indirect ELISA methods for detection of dog antibodies against rabies virus based on the recombinant matrix protein (M) and phosphoprotein (P). The M gene of rabies virus LEP-Flury strain was amplified by PCR and cloned into prokaryotic expression vector pGEX-6P-1. The resultant constructed pGEX-RV-M plasmid was transformed into BL21 (DE3) and protein expression was analyzed by SDS-PAGE and Western blot. The results of SDS-PAGE showed that the M protein was efficiently expressed, which were mainly soluble, and purified with the affinity chromatography. The results of Western blot indicated that the recombinant protein M showed good immunogenicity. The indirect ELISA method was established with the purified recombinant protein M, and a total of 95 serum samples were detected by the method and the indirect ELISA method based on the recombinant protein P respectively. The results showed that compared with the commercially available ELISA kit coating RV as antigen, the indirect ELISA method based on recombinant protein P had a higher coincidence rate, and it can replace the ELISA method based on RV.

CLC Number:

CHENG Chao-fei，GONG Miao-miao，TIAN Kang-le，WANG Yong，ZHAO Zhan-zhong，SHI Li-jun，LI Gang. Comparative Study on the Indirect ELISA Methods Based on the Different Recombinant Proteins of Rabies Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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Comparative Study on the Indirect ELISA Methods Based on the Different Recombinant Proteins of Rabies Virus

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