Prehaud C, Hellebrand E, Coudrier D, Volchkov V E, Volchkova V A, Feldmann H, Le Guenno B, Bouloy M
Unité des Arbovirus et virus des Fièvres Hémorragiques, Institut Pasteur, Paris, France.
J Gen Virol. 1998 Nov;79 ( Pt 11):2565-72. doi: 10.1099/0022-1317-79-11-2565.
After cloning and sequencing the glycoprotein (GP) gene of one of the Gabonese strains of Ebola virus isolated during the 1994-1996 outbreak, it was shown that the circulating virus was of the Zaire subtype. This was confirmed in this study by cloning and sequencing the nucleoprotein (NP) gene of this strain. These two structural proteins were also expressed as recombinant proteins and used in ELISA tests. NP was expressed as a His-tagged fusion protein in Escherichia coli and was purified on resins charged with nickel ions. GP was expressed by means of recombinant baculoviruses in Spodoptera frugiperda cells. Both recombinant proteins reacted positively in ELISAs for the detection of IgG antibodies in convalescent human sera from Gabon and Zaire. The difference in the relative titres of anti-NP and -GP antibodies was variable, depending on the sera. In addition, the recombinant NP reacted with heterologous sera from Côte d'Ivoire and was used successfully to detect IgM antibodies by mu-capture ELISA in sera from Gabonese patients.
在对1994 - 1996年埃博拉病毒爆发期间从加蓬分离出的一种毒株的糖蛋白（GP）基因进行克隆和测序后，结果显示流行病毒属于扎伊尔亚型。本研究通过对该毒株的核蛋白（NP）基因进行克隆和测序证实了这一点。这两种结构蛋白也被表达为重组蛋白并用于酶联免疫吸附测定（ELISA）试验。NP在大肠杆菌中表达为带His标签的融合蛋白，并在装有镍离子的树脂上进行纯化。GP通过重组杆状病毒在草地贪夜蛾细胞中表达。两种重组蛋白在用于检测来自加蓬和扎伊尔康复期人类血清中IgG抗体的ELISA试验中均呈阳性反应。抗NP和抗GP抗体的相对滴度差异因血清而异。此外，重组NP与来自科特迪瓦的异源血清发生反应，并成功用于通过μ捕获ELISA检测加蓬患者血清中的IgM抗体。
