Wen Gui-Ping, Tang Zi-Min, Yang Fan, Zhang Ke, Ji Wen-Fang, Cai Wei, Huang Shou-Jie, Wu Ting, Zhang Jun, Zheng Zi-Zheng, Xia Ning-Shao
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, and National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen, Fujian, People's Republic of China.
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, and National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen, Fujian, People's Republic of China
J Clin Microbiol. 2015 Mar;53(3):782-8. doi: 10.1128/JCM.01853-14. Epub 2014 Dec 24.
Hepatitis E virus (HEV) is a serious public health problem. The commonly used tests that are specific for current HEV infection diagnosis include the detection of anti-HEV IgM and HEV RNA. Here, we report an improved enzyme-linked immunosorbent assay (ELISA) method for HEV antigen detection with a linear range equivalent to 6.3 × 10(3) to 9.2 × 10(5) RNA copies per ml. The monoclonal antibody (MAb) 12F12, a high-ability MAb that binds HEV virus, was selected as the capture antibody from a panel of 95 MAbs. The positive period of HEV antigenemia in infected monkeys using this test was, on average, 3 weeks longer than previously reported and covered the majority of the acute phase. The positive detection rates of IgM, RNA, and new antigen from the first serum samples collected from 16 confirmed acute hepatitis E patients were 81% (13/16), 81% (13/16), and 100% (16/16), respectively. In three patients, the initial serum specimens that tested negative for IgM, despite the presence of symptoms of acute hepatitis and elevated alanine aminotransferase (ALT) levels, were positive for HEV antigen and HEV RNA. In contrast, the serum samples of the three RNA-negative patients were antigen positive (and IgM positive), possibly due to the degradation of HEV nucleic acids. Our results suggest that this new antigen detection method has acceptable concordance with RNA detection and could serve as an important tool for diagnosing acute hepatitis E.
戊型肝炎病毒(HEV)是一个严重的公共卫生问题。目前用于诊断当前HEV感染的常用特异性检测方法包括抗HEV IgM和HEV RNA的检测。在此,我们报告一种改进的酶联免疫吸附测定(ELISA)方法用于HEV抗原检测,其线性范围相当于每毫升6.3×10³至9.2×10⁵个RNA拷贝。单克隆抗体(MAb)12F12是一种结合HEV病毒能力很强的单克隆抗体,从95种单克隆抗体中被选为捕获抗体。使用该检测方法,感染猴子中HEV抗原血症的阳性期平均比先前报道的长3周,且涵盖了大部分急性期。从16例确诊的急性戊型肝炎患者收集的首份血清样本中,IgM、RNA和新抗原的阳性检出率分别为81%(13/16)、81%(13/16)和100%(16/16)。在3例患者中,尽管有急性肝炎症状且丙氨酸转氨酶(ALT)水平升高,但初始血清标本抗HEV IgM检测为阴性,而HEV抗原和HEV RNA检测为阳性。相反,3例RNA阴性患者的血清样本抗原呈阳性(且IgM呈阳性),这可能是由于HEV核酸降解所致。我们的结果表明,这种新的抗原检测方法与RNA检测具有可接受的一致性,可作为诊断急性戊型肝炎的重要工具。