Nagayama R, Tsuda F, Okamoto H, Wang Y, Mitsui T, Tanaka T, Miyakawa Y, Mayumi M
First Department of Internal Medicine, Teikyo University School of Medicine, Tokyo, Japan.
J Clin Invest. 1993 Sep;92(3):1529-33. doi: 10.1172/JCI116731.
Hepatitis C virus (HCV) samples in 155 sera, from patients with chronic non-A, non-B liver disease and blood donors, were grouped into four genotypes (I, II, III, and IV) by amplification of core-gene sequences by polymerase chain reaction with type-specific primers. HCV genotypes were compared with various HCV-associated antibodies detectable by the first-generation ELISA (ELISA-1) with C100-3 protein and a second-generation immunoblot assay with four recombinant HCV proteins. Antibodies to C100-3 protein and those to its subsequence (5-1-1) were detected in 13 (93%) and 12 (86%), respectively, of 14 sera with genotype I HCV; 56 (79%) and 58 (82%) of 71 sera with genotype II; 13 (34%) and 6 (16%) of 38 sera with genotype III; and 11 (34%) and 4 (13%) of 32 sera with genotype IV. Amino acid sequences of C100-3 of genotype I HCV are conserved by approximately 90% in genotype II, but only by approximately 75% in genotypes III and IV. The sensitivity of ELISA-1, therefore, would be influenced by heterogeneity in C100-3 sequences of different genotypes.
采用型特异性引物通过聚合酶链反应扩增核心基因序列,将155份慢性非甲非乙型肝病患者及献血者血清中的丙型肝炎病毒(HCV)样本分为四种基因型(I、II、III和IV)。将HCV基因型与通过第一代ELISA(ELISA - 1)检测C100 - 3蛋白以及通过第二代免疫印迹法检测四种重组HCV蛋白时可检测到的各种HCV相关抗体进行比较。在14份基因型I HCV血清中,分别有13份(93%)和12份(86%)检测到针对C100 - 3蛋白及其亚序列(5 - 1 - 1)的抗体;在71份基因型II血清中,分别有56份(79%)和58份(82%);在38份基因型III血清中,分别有13份(34%)和6份(16%);在32份基因型IV血清中,分别有11份(34%)和4份(13%)。基因型I HCV的C100 - 3氨基酸序列在基因型II中约90%保守,但在基因型III和IV中仅约75%保守。因此,ELISA - 1的敏感性会受到不同基因型C100 - 3序列异质性的影响。