不同戊型肝炎病毒(HEV)流行区病人及实验感染恒河猴血清能与病人粪便来源的HEV样病毒颗粒凝集,提示全球HEV流行有一主要型别。但迄今尚无可行的血清学研究方法。本研究中,自一HEV暴发病人粪便标本中提取RNA,合成cDNA后加接AB人工接头引物,在A链存在下,进行序列非依赖性单引物扩增。然后用EcoRI酶解,所得不同长度的cDNA克隆至噬菌体表达载体λgt11,构建cDNA文库。将这些cDNA克隆分别转染KM392菌,42℃培养5小时后转移至硝酸纤维滤膜。经HEV恢复期病人血清抗体检测,筛选出两个阳性克隆:406.4-2(M)及406.3-2(M)。将它们导入BNN103菌中,培养后得到两种β-半乳糖苷酶融合蛋白。经PAGE、Western印迹检测,发现两者均能与不同流行区急性和恢复期病人以及HEV感染恒河猴血清反应,提示这两种抗原 Agglutination of feces-derived HEV-like virus particles from patients with hepatitis E virus (HEV) endemic areas and experimental infected rhesus monkeys suggests a major type of global HEV epidemic. But so far there is no feasible serological research method. In this study, RNA was extracted from stool specimens from HEV-outbreak patients, followed by synthesis of cDNA followed by AB artificial linker primers, and sequence-independent single-primer amplification in the presence of A-chain. Then digested with EcoRI, and the resulting cDNAs of different lengths were cloned into the phage expression vector λgt11 to construct a cDNA library. These cDNA clones were transfected respectively with KM392 bacteria and cultured at 42 ° C for 5 hours and then transferred to a nitrocellulose filter. Two positive clones, 406.4-2 (M) and 406.3-2 (M), were screened out from HEV recovery patients’ serum antibodies. They were introduced into BNN103 bacteria and two β-galactosidase fusion proteins were obtained after culturing. By PAGE, Western blot test and found that both with different epidemic areas of acute and convalescent patients and HEV infected rhesus serum response, suggesting that these two antigens