Kaur M, Hyams K C, Purdy M A, Krawczynski K, Ching W M, Fry K E, Reyes G R, Bradley D W, Carl M
Accelerated Product Development and Infectious Disease Threat Assessment, Naval Medical Research Institute, Bethesda, MD 20814-5055.
Proc Natl Acad Sci U S A. 1992 May 1;89(9):3855-8. doi: 10.1073/pnas.89.9.3855.
Hepatitis E virus is responsible for both sporadic and epidemic hepatitis in developing countries. The nonenveloped virus is 27-34 nm in diameter and has been shown to contain a single-strand, positive-sense, polyadenylylated RNA genome of approximately 7.5 kilobases. The nucleotide sequence of the Burma strain of hepatitis E virus has been reported and three open reading frames (ORFs) have been identified. The deduced amino acid sequence from each of these ORFs was used to synthesize overlapping peptides (decamers overlapping at every fourth amino acid) on a solid phase. These peptides were then tested in an ELISA with pooled acute-phase sera from known cases of enterically transmitted non-A, non-B hepatitis collected in the Sudan. Linear B-cell epitopes were identified in all three ORFs. Epitopes were identified throughout the polyprotein encoded by ORF1, but they appeared to be particularly concentrated in the region of the RNA-dependent RNA polymerase. Distinct epitopes were identified in the presumed structural protein encoded by ORF2, and one epitope was identified close to the carboxyl terminus of the protein encoded by ORF3. These data precisely pinpoint linear B-cell epitopes recognized by antibodies from patients with acute hepatitis E and identify an antibody response directed against the RNA-dependent RNA polymerase.
戊型肝炎病毒在发展中国家可引发散发性和流行性肝炎。这种无包膜病毒直径为27 - 34纳米,已证实其含有一个约7.5千碱基的单链、正义、多聚腺苷酸化的RNA基因组。戊型肝炎病毒缅甸株的核苷酸序列已被报道,并且已鉴定出三个开放阅读框(ORF)。从这些ORF中的每一个推导的氨基酸序列用于在固相上合成重叠肽(每隔四个氨基酸重叠的十肽)。然后,这些肽在酶联免疫吸附测定(ELISA)中用来自苏丹收集的已知肠道传播的非甲非乙型肝炎病例的急性期血清池进行检测。在所有三个ORF中都鉴定出线性B细胞表位。表位在由ORF1编码的多蛋白中均有发现,但它们似乎特别集中在RNA依赖性RNA聚合酶区域。在由ORF2编码的假定结构蛋白中鉴定出不同的表位,并且在由ORF3编码的蛋白的羧基末端附近鉴定出一个表位。这些数据精确地确定了急性戊型肝炎患者抗体识别的线性B细胞表位,并确定了针对RNA依赖性RNA聚合酶的抗体反应。