Attallah Abdelfattah M, Ismail Hisham, Shiha Gamal E, Abou-Dobara Mohamed I, El-Sherbiny Rania E, El-Dosoky Ibrahim
Research & Development Department, Biotechnology Research Center, New Damietta, Egypt.
Clin Chim Acta. 2008 Feb;388(1-2):115-22. doi: 10.1016/j.cca.2007.10.019. Epub 2007 Oct 26.
The identification of native HCV antigens may prove very useful in the diagnosis and early treatment of HCV infection. Here, we aimed to identify and partially characterize a native HCV-NS4 antigen.
The western blot, ELISA and immunohistochemical staining were used to identify the native antigen in sera and liver biopsies of HCV serotype 4 infected patients.
The native NS4 antigen was identified in serum at 27-kDa molecular weight, in addition to a lower approximately 21-kDa antigen. The purified HCV antigen showed a polypeptide band at 27-kDa when analyzed by silver stained SDS-PAGE and a single peak at 7.6 min by capillary zone electrophoresis. The immunostaining pattern of hepatocytes was cytoplasmic with mainly coarse granular and diffuse pattern based on specific rabbit antisera to the native HCV antigen. A highly significant correlation (r=0.797, p<0.0001) was shown between serum concentrations of the HCV-NS4 antigen and HCV-RNA. Also, antigen detection rates were increased (p<0.05) with the progression of liver disease.
A native HCV-NS4 antigen was identified and partially characterized as 27-kDa protein and the NS4 antigenemia based ELISA test can serve as a useful addition to HCV diagnostic methods, especially under field condition.
鉴定天然丙型肝炎病毒（HCV）抗原可能在HCV感染的诊断和早期治疗中非常有用。在此，我们旨在鉴定并部分表征一种天然HCV-NS4抗原。
采用蛋白质印迹法、酶联免疫吸附测定法（ELISA）和免疫组织化学染色法，对HCV 4型感染患者的血清和肝活检组织中的天然抗原进行鉴定。
除了一种分子量约为21 kDa的较低抗原外，在血清中鉴定出分子量为27 kDa的天然NS4抗原。经银染十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析，纯化的HCV抗原显示出一条27 kDa的多肽带，经毛细管区带电泳分析，在7.6分钟处有一个单峰。基于针对天然HCV抗原的特异性兔抗血清，肝细胞的免疫染色模式为细胞质染色，主要为粗颗粒状和弥漫性模式。HCV-NS4抗原的血清浓度与HCV-RNA之间显示出高度显著的相关性（r = 0.797，p < 0.0001）。此外，随着肝病进展，抗原检测率增加（p < 0.05）。
鉴定出一种天然HCV-NS4抗原，并将其部分表征为27 kDa蛋白，基于NS4抗原血症的ELISA检测可作为HCV诊断方法的有益补充，尤其是在现场条件下。
